EP0786522A2 - Molécules de RNA enzymatiques pour le traitement de conditions sténotiques - Google Patents

Molécules de RNA enzymatiques pour le traitement de conditions sténotiques Download PDF

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EP0786522A2
EP0786522A2 EP97101534A EP97101534A EP0786522A2 EP 0786522 A2 EP0786522 A2 EP 0786522A2 EP 97101534 A EP97101534 A EP 97101534A EP 97101534 A EP97101534 A EP 97101534A EP 0786522 A2 EP0786522 A2 EP 0786522A2
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ribozyme
rna
rna molecule
ribozymes
enzymatic rna
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EP0786522A3 (fr
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Sean M. Sullivan
Kenneth G. Draper
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Sirna Therapeutics Inc
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Ribozyme Pharmaceuticals Inc
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Definitions

  • This invention relates to methods for inhibition of various animal diseases, including: a stenotic condition, in particular the inhibition of genetic expression which leads to the reduction or elimination of cellular proliferation in the areas of restenotic plaques.
  • Stenosis is the occurrence of a blockage in a blood vessel. Such blockages may lead to impairment of functions or even death, dependent upon which vessel it occurs in and its size. These may be prevented as described below.
  • Restenosis is a disease state which occurs as a sequelae to percutaneous transluminal angioplasty (PCTA) or coronary artery bypass grafting (CABG) treatments of cardiovascular disease.
  • PCTA percutaneous transluminal angioplasty
  • CABG coronary artery bypass grafting
  • Pathologic mechanisms associated with restenosis can be divided into three categories based upon when the restenosis occurs. The three categories and their mechanisms are 1) pre- and post-PCTA: thrombosis, platelet activation and thrombin generation; 2) immediate, after PCTA: elastic recoil; and 3) delayed, after PCTA: fibrocellular proliferation.
  • Acute or immediate restenosis occurs in approximately 10% of the patients undergoing PCTA and slower-developing restenosis (six month onset) occurs in approximately 30% of those patients.
  • the estimated number of PCTA patients in 1990 was 300,000 - 400,000. Thus, there are about 150,000 cases of restenosis per year in the United States.
  • the presently preferred chemotherapeutic treatment of patients is the use of streptokinase, urokinase or other thrombolytic compounds, such as fish oil, anticoagulants, ACE (angiotensin converting enzyme) inhibitors, aspirin and cholesterol lowering compounds; alternative treatment includes the surgical incorporation of endoluminal stents. It is reported that none of the current therapies have significantly impacted the rates of restenosis occurrence. A number of compounds are currently in preclinical evaluations. Platelet inhibitors include GR32191, Sultroban, Ketanserin, and fish oil.
  • Angiopeptin is being tested as a growth factor inhibitor and Lovostatin, Enoxaparin, RD Heparin, Cilazapril and Fosinopril are being investigated as smooth muscle cell proliferation inhibitors. While platelet inhibitors are being tested for prevention of restenosis, it appears that these compounds will not be efficacious as short-term treatments.
  • One of the biggest problems with current therapies is the occurrence of pharmacologic side-effects. These effects not only create other physiological problems, but also decrease the levels of patient compliance, thereby reducing the therapeutic efficacy of the treatments.
  • the proliferation of antherophils may be induced through a host of genetic activations, but the best candidate for targeting smooth muscle proliferation is the c-myb gene.
  • the c-myb protein binds DNA and activates DNA replication and cellular growth.
  • the role of c-myb in smooth muscle cell replication has been documented in bovine cells, and the expression of c-myb has been shown to activate cellular replication in chicken embryo fibroblasts and human T-lymphocytes.
  • the invention features use of ribozymes to treat or prevent various animal diseases, in particular, those human diseases noted above.
  • Stenosis is treated e.g. , by inhibiting the activation of smooth muscle proliferation by inhibiting the expression of the cellular c-myb gene. Cleavage of targeted mRNAs (c-myb mRNAs) expressed in endothelial cells and smooth muscle cells represses activation of cellular replication and abnormal proliferation of the smooth muscle or endothelial cells.
  • c-myb mRNAs targeted mRNAs
  • Ribozymes are RNA molecules having an enzymatic activity which is able to repeatedly cleave other separate RNA molecules in a nucleotide base sequence specific manner. It is alleged that such enzymatic RNA molecules can be targeted to virtually any RNA transcript and efficient cleavage has been achieved in vitro . Kim et al., 84 Proc. Natl. Acad. Sci. USA 8788, 1987; Haseloff and Gerlach, 334 Nature 585, 1988; Cech, 260 JAMA 3030, 1988; and Jefferies et al., 17 Nucleic Acids Research 1371, 1989.
  • Ribozymes act by first binding to a target RNA. Such binding occurs through the target RNA binding portion of a ribozyme which is held in close proximity to an enzymatic portion of the RNA which acts to cleave the target RNA. Thus, the ribozyme first recognizes and then binds a target RNA through complementary base-pairing, and once bound to the correct site, acts enzymatically to cut the target RNA. Strategic cleavage of such a target RNA will destroy its ability to direct synthesis of an encoded protein. After a ribozyme has bound and cleaved its RNA target it is released from that RNA to search for another target and can repeatedly bind and cleave new targets.
  • ribozyme The enzymatic nature of a ribozyme is advantageous over other technologies, such as antisense technology (where a nucleic acid molecule simply binds to a nucleic acid target to block its translation) since the effective concentration of ribozyme necessary to effect a therapeutic treatment is lower than that of an antisense oligonucleotide.
  • This advantage reflects the ability of the ribozyme to act enzymatically.
  • a single ribozyme molecule is able to cleave many molecules of target RNA.
  • the ribozyme is a highly specific inhibitor, with the specificity of inhibition depending not only on the base pairing mechanism of binding, but also on the mechanism by which the molecule inhibits the expression of the RNA to which it binds. That is, the inhibition is caused by cleavage of the RNA target and so specificity is defined as the ratio of the rate of cleavage of the targeted RNA over the rate of cleavage of non-targeted RNA. This cleavage mechanism is dependent upon factors additional to those involved in base pairing. Thus, it is thought that the specificity of action of a ribozyme is greater than that of antisense oligonucleotide binding the same RNA site.
  • This class of chemicals exhibits a high degree of specificity for cleavage of the intended target mRNA. Consequently, the ribozyme agent will only affect cells expressing that particular gene, and will not be toxic to normal tissues.
  • the invention can be used to treat or prevent (prophylactically) restenosis
  • the preferred administration protocol is in vivo administration to reduce the level of those genes and the encoded mRNAs noted above.
  • the invention features an enzymatic RNA molecule (or ribozyme) which cleaves mRNA associated with development or maintenance of a restenotic condition, e.g. , mRNA encoding c-myb, and in particular, those mRNA targets disclosed in Table 1.
  • a restenotic condition e.g. , mRNA encoding c-myb, and in particular, those mRNA targets disclosed in Table 1.
  • enzymatic RNA molecule an RNA molecule which has complementarity in a substrate binding region to a specified mRNA target, and also has an enzymatic activity which is active to specifically cleave that mRNA. That is, the enzymatic RNA molecule is able to intermolecularly cleave mRNA and thereby inactivate a target mRNA molecule. This complementarity functions to allow sufficient hybridization of the enzymatic RNA molecule to the target RNA to allow the cleavage to occur. One hundred percent complementarity is preferred, but complementarity as low as 50-75% may also be useful in this invention. For in vivo treatment, complementarity between 30 and 45 bases is preferred.
  • the enzymatic RNA molecule is formed in a hammerhead motif, but may also be formed in the motif of a hairpin, hepatitis delta virus, group I intron or RNaseP-like RNA (in association with an RNA guide sequence).
  • RNA molecules of this invention has a specific substrate binding site which is complementary to one or more of the target gene RNA regions, and that it have nucleotide sequences within or surrounding that substrate binding site which impart an RNA cleaving activity to the molecule.
  • the invention features a mammalian cell which includes an enzymatic RNA molecule as described above.
  • the mammalian cell is a human cell.
  • the invention features an expression vector which includes nucleic acid encoding an enzymatic RNA molecule described above, located in the vector, e.g. , in a manner which allows expression of that enzymatic RNA molecule within a mammalian cell.
  • the invention features a use of an enzymatic RNA molecule for the manufacture of a pharmaceutical composition for treatment of a disease noted above
  • the invention provides a class of chemical cleaving agents which exhibit a high degree of specificity for the mRNA causative of a stenotic or prestenotic condition.
  • a condition includes any measurable indication of susceptibility to cardiac problems, and thus includes predisposition to such conditions or cardiovascular disease.
  • Such enzymatic RNA molecules can be delivered exogenously or endogenously to infected cells.
  • the small size (less than 40 nucleotides, preferably between 32 and 36 nucleotides in length) of the molecule allows the cost of treatment to be reduced.
  • ribozyme delivered for any type of treatment reported to date is an in vitro transcript having a length of 142 nucleotides. Synthesis of ribozymes greater than 100 nucleotides in length is very difficult using automated methods, and the therapeutic cost of such molecules is prohibitive. Delivery of ribozymes by expression vectors is primarily feasible using only ex vivo treatments. This limits the utility of this approach.
  • an alternative approach uses smaller ribozyme motifs (e.g. , of the hammerhead structure, shown generally in Fig. 1) and exogenous delivery. The simple structure of these molecules also increases the ability of the ribozyme to invade targeted regions of the mRNA structure.
  • the enzymatic RNA molecules of this invention can be used to treat stenotic or prestenotic conditions. Such treatment can also be extended to other related genes in nonhuman primates. Affected animals can be treated at the time of disease risk or detection, or in a prophylactic manner. This timing of treatment will reduce the chance of further disease damage.
  • Ribozymes of this invention may be used as diagnostic tools to examine genetic drift and mutations within diseased cells.
  • the close relationship between ribozyme activity and the structure of the target RNA allows the detection of mutations in any region of the molecule which alters the base-pairing and three-dimensional structure of the target RNA.
  • multiple ribozymes described in this invention one may map nucleotide changes which are important to RNA structure and function in vitro , as well as in cells and tissues. Cleavage of target RNAs with ribozymes may be used to inhibit gene expression and define the role (essentially) of specified gene products in the progression of disease. In this manner, other genetic targets may be defined as important mediators of the disease.
  • Fig. 1 is a diagrammatic representation of a hammerhead motif ribozyme showing stems I, II and III (marked (I), (II) and (III) respectively) interacting with a target region. The 5' and 3' ends of both ribozyme and target are shown. Dashes indicate base-paired nucleotides.
  • Ribozymes targeting selected regions of mRNA associated with a selected disease are chosen to cleave the target RNA in a manner which preferably inhibits translation of the RNA. Genes are selected such that inhibition of translation will preferably inhibit cell replication, e.g. , by inhibiting production of a necessary protein. Selection of effective target sites within these critical regions of mRNA entails testing the accessibility of the target RNA to hybridization with various oligonucleotide probes. These studies can be performed using RNA probes and assaying accessibility by cleaving the hybrid molecule with RNaseH (see below).
  • ribozyme probes designed from secondary structure predictions of the mRNAs, and assaying cleavage products by polyacrylamide gel electrophoresis (PAGE), to detect the presence of cleaved and uncleaved molecules.
  • PAGE polyacrylamide gel electrophoresis
  • the method involves identifying potential cleavage sites for a hammerhead ribozyme, and then testing each of these sites to determine their suitability as targets by ensuring that secondary structure formation is minimal.
  • the mRNA sequences are compared in an appropriate target region. Putative ribozyme cleavage sites are found. These sites represent the preferable sites for hammerhead ribozyme cleavage within these target mRNAs.
  • RNA substrates corresponding to each of the mRNA sites are designed.
  • Each substrate is composed of two to three nucleotides at the 5' and 3' ends that will not base pair with a corresponding ribozyme recognition region.
  • the unpaired regions flanked a central region of 12-14 nucleotides to which complementary arms in the ribozyme are designed.
  • each substrate sequence is predicted using a standard PC fold computer program. Sequences which give a positive free energy of binding are accepted. Sequences which give a negative free energy are modified by trimming one or two bases from each of the ends. If the modified sequences are still predicted to have a strong secondary structure, they are rejected.
  • ribozymes are designed to each of the RNA substrates. Ribozyme folding is also analyzed using PC fold.
  • Ribozyme molecules are sought which form hammerhead motif stem II (see Fig. 1) regions and contain flanking arms which are devoid of intramolecular base pairing. Often the ribozymes are modified by trimming a base from the ends of the ribozyme, or by introducing additional base pairs in stem II to achieve the desired fold. Ribozymes with incorrect folding are rejected. After substrate/ribozyme pairs are found to contain correct intramolecular structures, the molecules are folded together to predict intermolecular interactions. A schematic representation of a ribozyme with its coordinate base pairing to its cognate target sequence is shown in Fig. 1.
  • ribonuclease H assay provides a quick test of the use of the target site without requiring synthesis of a ribozyme. It can be used to screen for sites most suited for ribozyme attack.
  • Ribozymes useful in this invention can be produced by gene transcription as described by Cech, supra , or by chemical synthesis. Chemical synthesis of RNA is similar to that for DNA synthesis. The additional 2'-OH group in RNA, however, requires a different protecting group strategy to deal with selective 3'-5' internucleotide bond formation, and with RNA susceptibility to degradation in the presence of bases. The decently developed method of RNA synthesis utilizing the t-butyldimethylsilyl group for the protection of the 2' hydroxyl is the most reliable method for synthesis of ribozymes. The method reproducibly yields RNA with the correct 3'-5' internucleotide linkages, with average coupling yields in excess of 99%, and requires only a two-step deprotection of the polymer.
  • a method, based upon H-phosphonate chemistry of phosphoroamidites gives a relatively lower coupling efficiency than a method based upon phosphoroamidite chemistry. This is a problem for synthesis of DNA as well.
  • a promising approach to scale-up of automatic oligonucleotide synthesis has been described recently for the H-phosphonates.
  • a combination of a proper coupling time and additional capping of "failure" sequences gave high yields in the synthesis of oligodeoxynucleotides in scales in the range of 14 ⁇ moles with as little as 2 equivalents of a monomer in the coupling step.
  • Another alternative approach is to use soluble polymeric supports (e.g. , polyethylene glycols), instead of the conventional solid supports. This method can yield short oligonucleotides in hundred milligram quantities per batch utilizing about 3 equivalents of a monomer in a coupling step.
  • ribozyme structure can be made to enhance the utility of ribozymes. Such modifications will enhance shelf-life, half-life in vitro , stability, and ease of introduction of such ribozymes to the target site, e.g. , to enhance penetration of cellular membranes, and confer the ability to recognize and bind to targeted cells.
  • Exogenous delivery of ribozymes benefits from chemical modification of the backbone, e.g. , by the overall negative charge of the ribozyme molecule being reduced to facilitate diffusion across the cell membrane.
  • the present strategies for reducing the oligonucleotide charge include: modification of internucleotide linkages by methylphosphonates, use of phosphoramidites, linking oligonucleotides to positively charged molecules, and creating complex packages composed of oligonucleotides, lipids and specific receptors or effectors for targeted cells. Examples of such modifications include sulfur-containing ribozymes containing phosphorothioates and phosphorodithioates as internucleotide linkages in RNA.
  • Ribozymes may also contain ribose modified ribonucleotides.
  • Pyrimidine analogues are prepared from uridine using a procedure employing diethylamino sulphur trifluoride (DAST) as a starting reagent.
  • Ribozymes can also be either electrostatically or covalently attached to polymeric cations for the purpose of reducing charge.
  • the polymer can be attached to the ribozyme by simply converting the 3'-end to a ribonucleoside dialdehyde which is obtained by a periodate cleavage of the terminal 2',3'-cis diol system.
  • other possible modifications may include different linker arms containing carboxyl, amino or thiol functionalities.
  • Yet further examples include use of methylphosphonates and 2'-O-methylribose and 5' or 3' capping or blocking with m 7 GpppG or m 3 2,2,7 GpppG.
  • a kinased ribozyme is contacted with guanosine triphosphate and guanyltransferase to add an m 3 G cap to the ribozyme.
  • the ribozyme can be gel purified using standard procedure. To ensure that the ribozyme has the desired activity, it may be tested with and without the 5' cap using standard procedures to assay both its enzymatic activity and its stability.
  • Synthetic ribozymes including those containing various modifiers, can be purified by high pressure liquid chromatography (HPLC).
  • HPLC high pressure liquid chromatography
  • Other liquid chromatography techniques, employing reverse phase columns and anion exchangers on silica and polymeric supports may also be used.
  • Synthesis is done on a 1 ⁇ mole scale using a 1 ⁇ mole RNA reaction column (Glen Research).
  • the average coupling efficiencies are between 97% and 98% for the 394 model, and between 97% and 99% for the 380B model, as determined by a calorimetric measurement of the released trityl cation.
  • Blocked ribozymes are cleaved from the solid support (e.g. , CPG), and the bases and diphosphoester moiety deprotected in a sterile vial by dry ethanolic ammonia (2 mL) at 55°C for 16 hours. The reaction mixture is cooled on dry ice. Later, the cold liquid is transferred into a sterile screw cap vial and lyophilized.
  • the solid support e.g. , CPG
  • dry ethanolic ammonia (2 mL) at 55°C for 16 hours.
  • the reaction mixture is cooled on dry ice. Later, the cold liquid is transferred into a sterile screw cap vial and lyophilized.
  • the residue is suspended in 1 M tetra-n-butylammonium fluoride in dry THF (TBAF), using a 20 fold excess of the reagent for every silyl group, for 16 hours at ambient temperature (about 15-25°C).
  • THF dry THF
  • the reaction is quenched by adding an equal volume of sterile 1 M triethylamine acetate, pH 6.5.
  • the sample is cooled and concentrated on a SpeedVac to half the initial volume.
  • the ribozymes are purified in two steps by HPLC on a C4 300 ⁇ 5 mm DeltaPak column in an acetonitrile gradient.
  • the first step is a separation of 5'-DMT-protected ribozyme(s) from failure sequences lacking a 5'-DMT group.
  • Solvents used for this step are: A (0.1 M triethylammonium acetate, pH 6.8) and B (acetonitrile).
  • the elution profile is: 20% B for 10 minutes, followed by a linear gradient of 20% B to 50% B over 50 minutes, 50% B for 10 minutes, a linear gradient of 50% B to 100% B over 10 minutes, and a linear gradient of 100% B to 0% B over 10 minutes.
  • the second step is a purification of a completely deblocked ribozyme by a treatment of 2% trifluoroacetic acid on a C4 300 ⁇ 5 mm DeltaPak column in an acetonitrile gradient.
  • Solvents used for this second step are: A (0.1 M triethylammonium acetate, pH 6.8) and B (80% acetonitrile, 0.1 M triethylammonium acetate, pH 6.8).
  • the elution profile is: 5% B for 5 minutes, a linear gradient of 5% B to 15% B over 60 minutes, 15% B for 10 minutes, and a linear gradient of 15% B to 0% B over 10 minutes.
  • the fraction containing ribozyme is cooled and lyophilized on a SpeedVac. Solid residue is dissolved in a minimum amount of ethanol and sodium perchlorate in acetone. The ribozyme is collected by centrifugation, washed three times with acetone, and lyophilized.
  • a promoter e.g. , the human cytomegalovirus immediate early promoter or human beta actin promoter
  • a promoter should be chosen to be a relatively strong promoter, and expressible both in vitro and in vivo (e.g. , the human cytomegalovirus immediate early promoter or human beta actin promoter).
  • Such a promoter should express the ribozyme at a level suitable to effect production of enough ribozyme to destroy a target RNA, but not at too high a level to prevent other cellular activities from occurring (unless cell death itself is desired).
  • a hairpin at the 5' end of the ribozyme is useful to ensure that the required transcription initiation sequence (GG or GGG or GGGAG) does not bind to some other part of the ribozyme and thus affect regulation of the transcription process.
  • the 5' hairpin is also useful to protect the ribozyme from 5'-3' exonucleases.
  • a selected hairpin at the 3' end of the ribozyme gene is useful since it acts as a transcription termination signal, and protects the ribozyme from 3'-5' exonuclease activity.
  • a known termination signal is that present on the T7 RNA polymerase system. This signal is about 30 nucleotides in length. Other 3' hairpins of shorter length can be used to provide good termination and RNA stability. Such hairpins can be inserted within the vector sequences to allow standard ribozymes to be placed in an appropriate orientation and expressed with such sequences attached.
  • Poly(A) tails are also useful to protect the 3' end of the ribozyme. These can be provided by either including a poly(A) signal site in the expression vector (to signal a cell to add the poly(A) tail in vivo ), or by introducing a poly(A) sequence directly into the expression vector. In the first approach the signal must be located to prevent unwanted secondary structure formation with other parts of the ribozyme. In the second approach, the poly(A) stretch may reduce in size over time when expressed in vivo , and thus the vector may need to be checked over time. Care must be taken in addition of a poly(A) tail which binds poly(A) binding proteins which prevent the ribozyme from acting.
  • ribozymes Once the desired ribozymes are selected, synthesized and purified, they are tested in kinetic and other experiments to determine their utility. An example of such a procedure is provided below.
  • Crude synthetic ribozyme (typically 350 ⁇ g at a time) is purified by separation on a 15% denaturing polyacrylamide gel (0.75 mm thick, 40 cm long) and visualized by UV shadowing. Once excised, gel slices containing full length ribozyme are soaked in 5 ml gel elution buffer (0.5 M NH 4 OAc, 1 mM EDTA) overnight with shaking at 4°C. The eluent is desalted over a C-18 matrix (Sep-Pak cartridges, Millipore, Milford, MA) and vacuum dried. The dried RNA is resuspended in 50-100 ⁇ l TE (TRIS 10 mM, EDTA 1 mM, pH 7.2).
  • a ribozyme may advantageously be synthesized in two or more portions. Each portion of a ribozyme will generally have only limited or no enzymatic activity, and the activity will increase substantially (by at least 5-10 fold) when all portions are ligated (or otherwise juxtaposed) together.
  • a specific example of hammerhead ribozyme synthesis is provided below.
  • the method involves synthesis of two (or more) shorter "half" ribozymes and ligation of them together using T4 RNA ligase.
  • T4 RNA ligase For example, to make a 34 mer ribozyme, two 17 mers are synthesized, one is phosphorylated, and both are gel purified. These purified 17 mers are then annealed to a DNA splint strand complementary to the two 17 mers. (Such a splint is not always necessary.) This DNA splint has a sequence designed to locate the two 17 mer portions with one end of each adjacent each other. The juxtaposed RNA molecules are then treated with T4 RNA ligase in the presence of ATP. The 34 mer RNA so formed is then HPLC purified.
  • a substrate solution is made 1X in assay buffer (75 mM Tris-HCl, pH 7.6; 0.1 mM EDTA, 10 mM MgCl 2 ) such that the concentration of substrate is less than 1 nM.
  • a ribozyme solution typically 20 nM is made 1X in assay buffer and four dilutions are made using 1X assay buffer. Fifteen ⁇ l of each ribozyme dilution ( i.e. , 20, 16, 12, 8 and 4 nM) is placed in a separate tube. These tubes and the substrate tube are pre-incubated at 37°C for at least five minutes.
  • the reaction is started by mixing 15 ⁇ l of substrate into each ribozyme tube by rapid pipetting (note that final ribozyme concentrations are 10, 8, 6, 4, 2 nM). Five ⁇ l aliquots are removed at 15 or 30 second intervals and quenched with 5 ⁇ l stop solution (95% formamide, 20 mM EDTA xylene cyanol, and bromphenol blue dyes). Following the final ribozyme time point, an aliquot of the remaining substrate is removed as a zero ribozyme control.
  • the samples are separated on either 15% or 20% polyacrylamide gels. Each gel is visualized and quantitated with an Ambis beta scanner (Ambis Systems, San Diego, CA).
  • the substrates are prepared by transcription using T7 RNA polymerase and defined templates containing a T7 promoter, and DNA encoding appropriate nucleotides of the target RNA.
  • the substrate solution is made 1X in assay buffer (75 mM Tris-HCl, pH 7.6; 0.1 mM EDTA; 10 mM MgCl 2 ) and contains 58 nanomolar concentration of the long RNA molecules.
  • the reaction is started by addition of gel purified ribozymes to 1 ⁇ M concentration. Aliquots are removed at 20, 40, 60, 80 and 100 minutes, then quenched by the addition of 5 ⁇ l stop solution. Cleavage products are separated using denaturing PAGE. The bands are visualized and quantitated with an Ambis beta scanner.
  • Lipid molecules are dissolved in a volatile organic solvent (CHCl 3 , methanol, diethylether, ethanol, etc.). The organic solvent is removed by evaporation.
  • the lipid is hydrated into suspension with 0.1x phosphate buffered saline (PBS), then freeze-thawed 3x using liquid nitrogen and incubation at room temperature.
  • PBS 0.1x phosphate buffered saline
  • the suspension is extruded sequentially through a 0.4 ⁇ m, 0.2 ⁇ m and 0.1 ⁇ m polycarbonate filters at maximum pressure of 800 psi.
  • the ribozyme is mixed with the extruded liposome suspension and lyophilized to dryness.
  • the lipid/ribozyme powder is rehydrated with water to one-tenth the original volume.
  • the suspension is diluted to the minimum volume required for extrusion (O.4 ml for 1.5 ml barrel and 1.5 ml for 10 ml barrel) with 1xPBS and re-extruded through 0.4 ⁇ m, 0.2 ⁇ m, 0.1 ⁇ m polycarbonate filters.
  • the liposome entrapped ribozyme is separated from untrapped ribozyme by gel filtration chromatography (SEPHAROSE CL-4B, BIOGEL A5M).
  • the liposome extractions are pooled and sterilized by filtration through a 0.2 ⁇ m filter.
  • the free ribozyme is pooled and recovered by ethanol precipitation.
  • the liposome concentration is determined by incorporation of a radioactive lipid.
  • the ribozyme concentration is determined by labeling with 32 P. Rossi et al., 1992, supra (and references cited therein) describe other methods suitable for preparation of liposomes.
  • liposomes composed of DPPG/DPPC/Cholesterol (in a ratio of: 50/17/33) gave a punctate pattern of fluorescence, while DOPE/Egg PC/Cholesterol (30/37/33) gave a diffuse, homogeneous pattern of fluorescence in the cytoplasm.
  • Cell fractionation showed that 80% of the entrapped contents from the DPPG/DPPC/Cholesterol formulation was localized in the membrane fraction, whereas the DOPE/Egg PC/Cholesterol formulation was localized in the cytoplasm. Further characterization of the latter formulation showed that after 3 hours, 70% of the fluorescence was cytoplasmic and 30% was in the membrane. After 24 hours, uptake had increased 5 fold and the liposome contents were distributed 50/50 between the cytoplasmic and membrane fractions.
  • Liposomes containing 15 ribozymes ( 32 P-labeled) targeted to the HSV ICP4 mRNA were prepared and incubated with the cells. After 24 hours, 25% of the liposome dose was taken up with approximately 60,000 liposomes per cell. Thirty percent of the delivered ribozyme was intact after 24 hours. Cell fractionation studies showed 40% of the intact ribozyme to be in the membrane fraction and 52% of the intact ribozyme to be in the cytoplasmic fraction.
  • the efficacy of action of a chosen ribozyme may be tested in vivo using standard procedures in transformed cells or animals which express the target mRNA.
  • the accumulation of target mRNA in cells or the cleavage of the RNA by ribozymes or RNaseH can be quantified using an RNase protection assay.
  • antisense riboprobes are transcribed from template DNA using T7 RNA polymerase (U.S. Biochemical) in 20 ⁇ l reactions containing 1X transcription buffer (supplied by the manufacturer), 0.2 mM ATP, GTP and UTP, 1 U/ ⁇ l pancreatic RNase inhibitor (Boehringer Mannheim Biochemicals) and 200 ⁇ Ci 32 P-labeled CTP (800 Ci/mmol, New England Nuclear) for 1 hour at 37°C.
  • Template DNA is digested with 1 U RNase-free DNaseI (U.S. Biochemical, Cleveland, OH) at 37°C for 15 minutes and unincorporated nucleotides removed by G-50 SEPHADEX spin chromatography.
  • the target RNA can be transcribed in vitro using a suitable DNA template.
  • the transcript is purified by standard methods and digested with ribozyme at 37°C according to methods described later.
  • afflicted (mRNA-expressing) cells are harvested into 1 ml of PBS, transferred to a 1.5 ml EPPENDORF tube, pelleted for 30 seconds at low speed in a microcentrifuge, and lysed in 70 ⁇ l of hybridization buffer (4 M guanidine isothiocyanate, 0.1% sarcosyl, 25 mM sodium citrate, pH 7.5).
  • Cell lysate 45 ⁇ l or defined amounts of in vitro transcript (also in hybridization buffer) is then combined with 5 ⁇ l of hybridization buffer containing 5 x 10 5 cpm of each antisense riboprobe in 0.5 ml Eppendorf tubes, overlaid with 25 ⁇ l mineral oil, and hybridization accomplished by heating overnight at 55°C.
  • hybridization reactions are diluted into 0.5 ml RNase solution (20 U/ml RNaseA, 2 U/ml RNaseT1, 10 U/ml RNase-free DNaseI in 0.4 M NaCl), heated for 30 minutes at 37°C, and 10 ⁇ l of 20% SDS and 10 ⁇ l of Proteinase K (10 mg/ml) added, followed by an additional 30 minutes incubation at 37°C.
  • Hybrids are partially purified by extraction with 0.5 ml of a 1:1 mixture of phenol/chloroform; aqueous phases are combined with 0.5 ml isopropanol, and RNase-resistant hybrids pelleted for 10 minutes at room temperature (about 20°C) in a microcentrifuge.
  • Pellets are dissolved in 10 ⁇ l loading buffer (95% formamide, 1X TBE, 0.1% bromophenol blue, 0.1% xylene cylanol), heated to 95°C for five minutes, cooled on ice, and analyzed on 4% polyacrylamide/7 M urea gels under denaturing conditions.
  • the chosen ribozyme can be tested to determine its stability, and thus its potential utility. Such a test can also be used to determine the effect of various chemical modifications (e.g. , addition of a poly(A) tail) on the ribozyme stability and thus aid selection of a more stable ribozyme.
  • a reaction mixture contains 1 to 5 pmoles of 5' (kinased) and/or 3' labeled ribozyme, 15 ⁇ g of cytosolic extract and 2.5 mM MgCl 2 in a total volume of 100 ⁇ l. The reaction is incubated at 37°C.
  • a 3'-labeled ribozyme can be formed by incorporation of the 32 P-labeled cordycepin at the 3' OH using poly(A) polymerase.
  • the poly(A) polymerase reaction contains 40 mM Tris, pH 8, 10 mM MgCl 2 , 250 mM NaCl, 2.5 mM MnCl 2 ,; 3 ⁇ l 32 P cordycepin, 500 Ci/mM; and 6 units poly(A) polymerase in a total volume of 50 ⁇ l. The reaction mixture is incubated for 30 minutes at 37°C.
  • minor base changes can be made in the substrate cleavage region recognized by a specific ribozyme.
  • the substrate sequences can be changed at the central "C" nucleotide, changing the cleavage site from a GUC to a GUA motif.
  • the K cat /K m values for cleavage using each substrate are then analyzed to determine if such a change increases ribozyme cleavage rates. Similar experiments can be performed to address the effects of changing bases complementary to the ribozyme binding arms. Changes predicted to maintain strong binding to the complementary substrate are preferred.
  • cleavage rates of ribozymes containing varied arm lengths, but targeted to the same length of short RNA substrate can be tested. Minimal arm lengths are required and effective cleavage varies with ribozyme/substrate combinations.
  • the cleavage activity of selected ribozymes can be assessed using target mRNA substrates.
  • the assays are performed in ribozyme excess and approximate K cat /K min values obtained. Comparison of values obtained with short and long substrates indicates utility in vivo of a ribozyme.
  • Ribozymes are 32 P-end labeled, entrapped in liposomes and delivered to target mRNA-containing cells for three hours.
  • the cells are fractionated and ribozyme is purified by phenol/chloroform extraction.
  • cells (1x10 7 , T-175 flask) are scraped from the surface of the flask and washed twice with cold PBS.
  • the cells are homogenized by douncing 35 times in 4 ml of TSE (10 mM Tris, pH 7.4, 0.25 M Sucrose, mM EDTA). Nuclei are pelleted at 100xg for 10 minutes.
  • Subcellular organelles are pelleted at 200,000xg for two hours using an SW60 rotor.
  • the pellet is resuspended in 1 ml of H buffer (0.25 M Sucrose, 50 mM HEPES, pH 7.4).
  • the supernatant contains the cytoplasmic fraction (in approximately 3.7 ml).
  • the nuclear pellet is resuspended in 1 ml of 65% sucrose in TM (50 mM Tris, pH 7.4, 2.5 mM MgCl 2 ) and banded on a sucrose step gradient (1 ml nuclei in 65% sucrose TM, 1 ml 60% sucrose TM, 1 ml 55% sucrose TM, 50% sucrose TM, 300 ⁇ l 25% sucrose TM) for one hour at 37,000xg with an SW60 rotor.
  • the nuclear band is harvested and diluted to 10% sucrose with TM buffer. Nuclei are pelleted at 37,000xg using an SW60 rotor for 15 minutes and the pellet resuspended in 1 ml of TM buffer. Aliquots are size fractionated on denaturing polyacrylamide gels and the intracellular localization determined. By comparison to the migration rate of newly synthesized ribozyme, the various fractions containing intact ribozyme can be determined.
  • the cells may be fractioned as above and the purity of each fraction assessed by assaying enzyme activity known to exist in that fraction.
  • Ribozyme molecules may be synthesized with 5 phosphorothioate (ps), or 2'-Omethyl (2'-OMe) modifications at each end of the molecule. These molecules and a phosphodiester version of the ribozyme are end-labeled with 32 P and ATP using T4 polynucleotide kinase. Equal concentrations are added to the cell cytoplasmic extracts and aliquots of each taken at 10 minute intervals. The samples are size fractionated by denaturing PAGE and relative rates of nuclease resistance analyzed by scanning the gel with an Ambis ⁇ -scanner.
  • ribozymes are digested by the cytoplasmic extract, and which versions are relatively more nuclease resistant. Modified ribozymes generally maintain 80-90% of the catalytic activity of the native ribozyme when short RNA substrates are employed.
  • Unlabeled, 5' end-labeled or 3' end-labeled ribozymes can be used in the assays. These experiments can also be performed with human cell extracts to verify the observations.
  • Vero or HeLa cells were grown to 90-95% confluency in 175 cm 2 tissue culture flasks, scraped into 10 ml of cold phosphate buffered saline (PBS), then washed once in 10 ml of cold PBS and once in 10 ml of cold TSE (10 mM Tris, pH 7.4; 0.25 M sucrose; 1 mM EDTA). The cell pellets were resuspended in 4 ml of TSE, dounced 35x on ice, and the released nuclei pelleted by centrifugation at 1000g for 10 minutes.
  • PBS cold phosphate buffered saline
  • the nuclear pellet was resuspended in 1 ml of 65% sucrose TM (50 mM Tris, pH 7.4; 2.5 mM MgCl 2 ) and transferred to Beckman ultra-clear tubes.
  • sucrose TM solutions were layered on top of the sample: 1 ml 60%, 1 ml 55%, and 25% sucrose to the top of the tube. Gradients were spun in an SW60 rotor at 37,000g for 1 hour. HeLa nuclei banded at the 55-60% sucrose boundary and Vero nuclei banded at the 50-55% sucrose boundary. Nuclear bands were harvested, diluted to 10% sucrose with TM buffer, and pelleted by centrifugation at 37,000g for 15 minutes using an SW60 rotor.
  • the nuclear pellet was resuspended in 1 ml of TM buffer. Subcellular organelles and membrane components in the post nuclear supernatant were separated from the cytoplasmic fraction by centrifugation at 200,000g for 2 hours in an SW60 rotor. The pellet contained the membrane fraction, which was resuspended in 1 ml of H buffer (0.25 M sucrose; 50 mM HEPES, pH 7.4), and the supernatant contained the cytoplasmic fraction.
  • the reaction mixture contained 0.3 mg/ml 4-methylumbelliferyl-N-acetyl-glucosaminide; 20 mM sodium citrate; pH 4.5; 0.01% Triton X-100; and 100 ⁇ l of sample in a final volume of 500 ⁇ l (Harding et a]., 64 Cell 393, 1991).
  • the reactions were incubated at 37°C for 1 hour and stopped by the addition of 1.5 ml of stop buffer (0.13 M glycine, 0.07 M NaCl, 0.08 M sodium carbonate, pH 10.6).
  • the reaction product was quantitated in a Hitachi F-4010 fluorescence spectrophotometer by excitation of the fluorophore at 360 nm and analysis of the emission at 448 nm.
  • the assay medium contained 25 mM CAPS (3-(Cyclohexylamino)propanesulfonic acid), pH 10.6; 0.05% Triton X-100; 15 mM MgCl 2 ; 1.25 mg/ml Thymidine-5'-monophosphate-p-nitrophenyl ester; and 100 ⁇ l of sample in a total reaction volume of 200 ⁇ l.
  • the reactions were incubated at 37°C for 2 hours, then diluted to 1 ml with H 2 O and the absorbance was measured at 400 nm (Razell and Khorana, 234 J. Biol. Chem. 739, 1959).
  • the reaction contained 85 nM sodium citrate, pH 5.9; 0.12% Triton X-100; 0.1% sodium taurocholate; 5 mM 4-methylumbelliferyl ⁇ -D-glucopyranoside; and 125 ⁇ l of sample in a total volume of 250 ⁇ l (Kennedy and Cooper, 252 Biochem. J. 739, 1988).
  • the reaction was incubated at 37°C for 1 hour and stopped by the addition of 0.75 ml of stop buffer.
  • Product formation was measured in a fluorescence spectrophotometer by using an excitation wavelength of 360 nm and analysis of the emission at 448 nm.
  • the cytoplasmic enzyme marker, lactate dehydrogenase was assayed in an assay mixture containing 0.2 M Tris; pH 7.4; 0.22 mM NADH; 1 mM sodium pyruvate; and 50 ⁇ l of sample in a final volume of 1.05 ml. Enzyme levels were determined by decreased absorbency at 350 nm resulting from the oxidation of NADH at room temperature (Silverstein and Boyer, 239 J. Biol. Chem. 3901, 1964).
  • Lactate dehydrogenase was found predominantly in the cytoplasmic fractions of both Vero and HeLa cells, while ⁇ -glucocerebrosidase and alkaline phosphodiesterase were found almost exclusively in the membranous fractions.
  • the hexosaminidase activity in Vero cell fractions was concentrated in the membranous fraction (70%) with about 20% in the cytoplasmic fraction.
  • the isolation of enzyme markers with the appropriate cellular compartment demonstrated that cytoplasmic, membranous and nuclear fractions can be isolated with minimal intercompartmental contamination using this fractionation scheme.
  • the basic oligonucleotide digestion reaction contained substrate nucleic acid (an RNA oligonucleotide of 36 nucleotides) and cell fraction extract in a total volume of 100 ⁇ l. Aliquots (7 ⁇ l) were taken after various periods of incubation at 37°C and added to 7 ⁇ l of gel loading buffer (95% formamide, 0.1% bromophenol blue, 0.1% xylene cyanol, and 20 mM EDTA). The samples were separated by electrophoresis on a 7 M urea, 20% polyacrylamide gel.
  • substrate nucleic acid an RNA oligonucleotide of 36 nucleotides
  • cell fraction extract in a total volume of 100 ⁇ l. Aliquots (7 ⁇ l) were taken after various periods of incubation at 37°C and added to 7 ⁇ l of gel loading buffer (95% formamide, 0.1% bromophenol blue, 0.1% xylene cyanol, and 20 mM EDTA). The samples were separated by
  • Intact ribozymes were visualized either by staining with Stains-all (United States Biochemical, Cleveland, OH), or autoradiography of 32 P-labeled ribozyme. The stained gels and X-ray films were scanned on a Bio 5000 density scanner (U.S. Biochemical). Ribozymes were 5' end-labeled with T4 polynucleotide kinase (U.S. Biochemical) using 10 ⁇ Ci of 32 P ⁇ -ATP (3,000 Ci/mmole, New England Nuclear, Boston, MA), and 20-25 pmoles of ribozymes. The unincorporated nucleotides were separated from the product by G-50 spin chromatography.
  • Nuclease assays contained 1-2 pmoles of 32 P-labeled ribozyme. All oligonucleotides were synthesized on an Applied Biosystems 394 DNA/RNA synthesizer (Applied Biosystems Inc., Foster City, CA) according to manufacturer's protocols. The nuclear fractions were resuspended in a buffer containing 2.5 mM MgCl 2 . Experiments involving the nuclear fractions were performed in the presence of 1 mM Mg +2 , or in combination with 1 mM Mn +2 , Ca +2 , or Zn +2 .
  • RNA probes were produced from PCR-amplified template DNA using T7 RNA polymerase (U.S. Biochemical) in the presence of 32 P ⁇ -CTP (3,000 Ci/mmole, New England Nuclear, Boston, MA). Template DNA was inactivated with 1 unit of RNase-free DNaseI for 15 minutes at 37°C.
  • Hybridization reactions were covered with mineral oil and incubated at 55°C for 12-16 hours, after which the hybridization reaction was mixed with 500 ⁇ l of RNase buffer (0.4 M NaCl, 20 ⁇ g/ml RNaseA, 2 units/ml T1 RNase) and incubated for 30 minutes at 37°C. RNase activity was quenched by incubation with 10 ⁇ l of 20% SDS and 10 ⁇ l of proteinase K (20 mg/ml), and the RNA was extracted using a phenol/chloroform mixture. The protected RNA fragment was purified by precipitation with an equal volume of isopropanol in the presence of 20 ⁇ g of carrier yeast tRNA.
  • RNase buffer 0.4 M NaCl, 20 ⁇ g/ml RNaseA, 2 units/ml T1 RNase
  • RNA pellets were resuspended in gel loading buffer, heated to 95°C for 5 minutes and separated by electrophoresis on a 5% polyacrylamide, 7 M urea gel. Protected fragments were visualized by autoradiography, and the films were scanned with a Bio 5000 density scanner.
  • ribozymes were incubated in various Vero and HeLa cellular fractions. Incubation of ribozymes in either membranous or nuclear fractions resulted in a linear decrease of intact molecules over time. In contrast, no digestion of ribozymes occurred during a 24 hour incubation in Vero cytoplasmic extracts, and HeLa cytoplasmic extracts exhibited a 20-30 minute delay in the onset of RNA digestion. After this refractory period, the rate of digestion was linear but not as rapid as the rates observed in any of the nuclear or membranous fractions.
  • the effect of four divalent cations (Mg +2 , Mn +2 , Ca +2 , and Zn +2 ) on the nuclease activity of the cellular fractions was assessed. Vero cytoplasmic extracts were stimulated by the addition of 1 mM Mg +2 or Mn +2 , while Ca +2 or Zn +2 had no effect. Nuclease activity in HeLa cytoplasmic extracts was enhanced only by the addition of 1 mM Zn +2 .
  • Vero and HeLa membranous fractions exhibited maximum nuclease activity with the addition of Mg +2 or Mn +2 ions, while the addition of Ca +2 significantly reduced activity of the HeLa membranous fraction and abolished nuclease activity in the Vero membranous fraction.
  • Addition of Zn +2 to both membranous fractions resulted in a loss of all RNase activity.
  • the Vero nuclear extract demonstrated roughly equivalent nuclease activity in the presence of either Mg +2 alone or a Mg +2 and Mn +2 ion combination, less in the presence of Mg +2 and Ca +2 , and no activity in the presence of Mg +2 and Zn +2 .
  • nuclease assays were performed using 1 mM Mg +2 in the presence and absence of 20 mM EDTA.
  • the Mg +2 was replaced with 1 mM Zn +2 .
  • the presence of 20 mM EDTA completely abolished nuclease activity in the Vero and HeLa cytoplasmic fractions and Vero nuclear fractions.
  • Nuclease activity in the HeLa membranous and nuclear fractions was partially inhibited by the addition of EDTA, while EDTA had no effect on the nuclease activity in the Vero membranous fraction.
  • reactions using DNA oligonucleotides were performed using different Vero fractions. All DNase activity in Vero cytoplasmic, membranous, and nuclear fractions was inhibited by 20 mM EDTA.
  • RNA oligonucleotides and HSV-1 mRNA were compared in the presence and absence of activity-enhancing divalent cations (1 mM Mg +2 , Vero cells; 1 mM Zn +2 , HeLa cells).
  • Total cellular RNA from HSV-1 infected Vero cells (8 mg) and tracer amounts of 32 P-5'-end-labeled RNA oligonucleotides (1 pmole) were incubated with Vero or HeLa cytoplasmic extracts. In the absence of divalent cations, no substantial decrease of intact ribozymes was detected in assays, although mRNA was digested in both Vero and HeLa cytoplasmic extracts.
  • hammerhead ribozymes were compared in both Vero and HeLa cell cytoplasmic, membranous and nuclear fractions.
  • Vero cytoplasmic and nuclear fractions were found to require Mg +2 for optimal nuclease activity, while the membranous fraction was not altered by the addition of divalent cations.
  • HeLa membranous and nuclear fractions were also activated by Mg +2 , while the cytoplasmic fractions required Zn +2 for nuclease activation.
  • Relative stabilities of ribozymes and mRNAs were compared in Vero and HeLa cytoplasmic fractions.
  • Selected ribozymes can be administered prophylactically, or to patients having disease conditions, e.g. , by exogenous delivery of the ribozyme to a desired tissue by means of an appropriate delivery vehicle, e.g. , a liposome, a controlled release vehicle, by use of iontophoresis, electroporation or ion paired molecules, or covalently attached adducts, and other pharmacologically approved methods of delivery.
  • routes of administration include intramuscular, aerosol, oral (tablet or pill form), topical, systemic, ocular, intraperitoneal and/or intrathecal.
  • Expression vectors for immunization with ribozymes and/or delivery of ribozymes are also suitable.
  • the specific delivery route of any selected ribozyme will depend on the use of the ribozyme. Generally, a specific delivery program for each ribozyme will focus on unmodified ribozyme uptake with regard to intracellular localization, followed by demonstration of efficacy. Alternatively, delivery to these same cells in an organ or tissue of an animal can be pursued. Uptake studies will include uptake assays to evaluate cellular ribozyme uptake, regardless of the delivery vehicle or strategy. Such assays will also determine the intracellular localization of the ribozyme following uptake, ultimately establishing the requirements for maintenance of steady-state concentrations within the cellular compartment containing the target sequence (nucleus and/or cytoplasm). Efficacy and cytotoxicity can then be tested. Toxicity will not only include cell viability but also cell function.
  • Some methods of delivery that may be used include:
  • ribozyme modifications like most small molecules, are taken up by cells, albeit slowly.
  • the ribozyme may be modified essentially at random, in ways which reduce its charge but maintains specific functional groups. This results in a molecule which is able to diffuse across the cell membrane, thus removing the permeability barrier.
  • ribozymes Modification of ribozymes to reduce charge is just one approach to enhance the cellular uptake of these larger molecules.
  • the random approach is not advisable since ribozymes are structurally and functionally more complex than small drug molecules.
  • the structural requirements necessary to maintain ribozyme catalytic activity are well understood by those in the art. These requirements are taken into consideration when designing modifications to enhance cellular delivery. The modifications are also designed to reduce susceptibility to nuclease degradation. Both of these characteristics should greatly improve the efficacy of the ribozyme.
  • Cellular uptake can be increased by several orders of magnitude without having to alter the phosphodiester linkages necessary for ribozyme cleavage activity.
  • Drug delivery vehicles are effective for both systemic and topical administration. They can be designed to serve as a slow release reservoir, or to deliver their contents directly to the target cell.
  • An advantage of using direct delivery drug vehicles is that multiple molecules are delivered per uptake. Such vehicles have been shown to increase the circulation half-life of drugs which would otherwise be rapidly cleared from the blood stream.
  • Some examples of such specialized drug delivery vehicles which fall into this category are liposomes, hydrogels, cyclodextrins, biodegradable nanocapsules, and bioadhesive microspheres.
  • liposomes are preferred. Liposomes increase intracellular stability, increase uptake efficiency and improve biological activity.
  • Liposomes are hollow spherical vesicles composed of lipids arranged in a similar fashion as those lipids which make up the cell membrane. They have an internal aqueous space for entrapping water soluble compounds and range in size from 0.05 to several microns in diameter. Several studies have shown that liposomes can deliver RNA to cells and that the RNA remains biologically active.
  • Lipofectin a liposome delivery vehicle originally designed as a research tool, Lipofectin, has been shown to deliver intact mRNA molecules to cells yielding production of the corresponding protein.
  • Liposomes offer several advantages: They are non-toxic and biodegradable in composition; they display long circulation half-lives; and recognition molecules can be readily attached to their surface for targeting to tissues. Finally, cost effective manufacture of liposome-based pharmaceuticals, either in a liquid suspension or lyophilized product, has demonstrated the viability of this technology as an acceptable drug delivery system.
  • Nonoparticles and hydrogels may be potential delivery vehicles for a ribozyme.
  • These carriers have been developed for chemotherapeutic agents and protein-based pharmaceuticals, and consequently, can be adapted for ribozyme delivery.
  • Topical administration of ribozymes is advantageous since it allows localized concentration at the site of administration with minimal systemic adsorption. This simplifies the delivery strategy of the ribozyme to the disease site and reduces the extent of toxicological characterization. Furthermore, the amount of material to be applied is far less than that required for other administration routes. Effective delivery requires the ribozyme to diffuse into the infected cells. Chemical modification of the ribozyme to neutralize negative charge may be all that is required for penetration. However, in the event that charge neutralization is insufficient, the modified ribozyme can be co-formulated with permeability enhancers, such as Azone or oleic acid, in a liposome.
  • permeability enhancers such as Azone or oleic acid
  • the liposomes can either represent a slow release presentation vehicle in which the modified ribozyme and permeability enhancer transfer from the liposome into the infected cell, or the liposome phospholipids can participate directly with the modified ribozyme and permeability enhancer in facilitating cellular delivery.
  • both the ribozyme and permeability enhancer can be formulated into a suppository formulation for slow release.
  • Ribozymes may also be systemically administered.
  • Systemic absorption refers to the accumulation of drugs in the blood stream followed by distribution throughout the entire body.
  • Administration routes which lead to systemic absorption include: intravenous, subcutaneous, intraperitoneal, intranasal, intrathecal and ophthalmic. Each of these administration routes expose the ribozyme to an accessible diseased tissue.
  • Subcutaneous administration drains into a localized lymph node which proceeds through the lymphatic network into the circulation. The rate of entry into the circulation has been shown to be a function of molecular weight or size.
  • the use of a liposome or other drug carrier localizes the ribozyme at the lymph node.
  • the ribozyme can be modified to diffuse into the cell, or the liposome can directly participate in the delivery of either the unmodified or modified ribozyme to the cell.
  • a liposome formulation containing phosphatidylethanolomidomethylthiosuccinimide which can deliver oligonucleotides to lymphocytes and macrophages is useful for certain conditions. Furthermore, a 200 nm diameter liposome of this composition was internalized as well as 100 nm diameter liposomes. The 200 nm liposomes exhibit a ten-fold greater packaging capacity than the 100 nm liposomes and can accommodate larger molecules such as a ribozyme expression vector. This ribozyme delivery system prevents mRNA expression in afflicted primary immune cells.
  • Whole blood studies show that the formulation is taken up by 90% of the lymphocytes after 8 hours at 37°C. Preliminary biodistribution and pharmacokinetic studies yielded 70% of the injected dose/gm of tissue in the spleen after one hour following intravenous administration.
  • Liposomes injected intravenously show accumulation in the liver, lung and spleen.
  • the composition and size can be adjusted so that this accumulation represents 30% to 40% of the injected dose.
  • the remaining dose circulates in the blood stream for up to 24 hours.
  • the chosen method of delivery should result in cytoplasmic accumulation and molecules should have some nuclease-resistance for optimal dosing.
  • Nuclear delivery may be used but is less preferable.
  • Most preferred delivery methods include liposomes (10-400 nm), hydrogels, controlled-release polymers, microinjection or electroporation (for ex vivo treatments) and other pharmaceutically applicable vehicles.
  • the dosage will depend upon the disease indication and the route of administration but should be between 100-200 mg/kg of body weight/day.
  • the duration of treatment will extend through the course of the disease symptoms, possibly continuously.
  • the number of doses will depend upon disease delivery vehicle and efficacy data from clinical trials.
  • ribozyme within the cell is dependent upon the rate of uptake and degradation. Decreasing the degree of degradation will prolong the intracellular half-life of the ribozyme.
  • chemically modified ribozymes e.g. , with modification of the phosphate backbone, or capping of the 5' and 3' ends of the ribozyme with nucleotide analogs may require different dosaging. Descriptions of useful systems are provided in the art cited above, all of which is hereby incorporated by reference herein.

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US889593A 1993-01-19 1993-01-19
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012030901A1 (fr) * 2010-08-31 2012-03-08 Novartis Ag Petits liposomes destinés à l'administration d'un arn codant pour un immunogène
US11291682B2 (en) 2010-07-06 2022-04-05 Glaxosmithkline Biologicals Sa Delivery of RNA to trigger multiple immune pathways
US11291635B2 (en) 2010-07-06 2022-04-05 Glaxosmithkline Biological Sa Virion-like delivery particles for self-replicating RNA molecules
US20220125723A1 (en) 2010-07-06 2022-04-28 Glaxosmithkline Biologicals Sa Lipid formulations with viral immunogens
US11639370B2 (en) 2010-10-11 2023-05-02 Glaxosmithkline Biologicals Sa Antigen delivery platforms
US11655475B2 (en) 2010-07-06 2023-05-23 Glaxosmithkline Biologicals Sa Immunisation of large mammals with low doses of RNA
US11759422B2 (en) 2010-08-31 2023-09-19 Glaxosmithkline Biologicals Sa Pegylated liposomes for delivery of immunogen-encoding RNA
US11896636B2 (en) 2011-07-06 2024-02-13 Glaxosmithkline Biologicals Sa Immunogenic combination compositions and uses thereof

Families Citing this family (544)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6469158B1 (en) 1992-05-14 2002-10-22 Ribozyme Pharmaceuticals, Incorporated Synthesis, deprotection, analysis and purification of RNA and ribozymes
US5804683A (en) * 1992-05-14 1998-09-08 Ribozyme Pharmaceuticals, Inc. Deprotection of RNA with alkylamine
US5686599A (en) * 1992-05-14 1997-11-11 Ribozyme Pharmaceuticals, Inc. Synthesis, deprotection, analysis and purification of RNA and ribozymes
US5977343A (en) 1992-05-14 1999-11-02 Ribozyme Pharmaceuticals, Inc. Synthesis, deprotection, analysis and purification of RNA and ribozymes
US5646042A (en) * 1992-08-26 1997-07-08 Ribozyme Pharmaceuticals, Inc. C-myb targeted ribozymes
US5891684A (en) * 1992-10-15 1999-04-06 Ribozyme Pharmaceuticals, Inc. Base-modified enzymatic nucleic acid
US5658780A (en) 1992-12-07 1997-08-19 Ribozyme Pharmaceuticals, Inc. Rel a targeted ribozymes
US5612215A (en) * 1992-12-07 1997-03-18 Ribozyme Pharmaceuticals, Inc. Stromelysin targeted ribozymes
US5811300A (en) * 1992-12-07 1998-09-22 Ribozyme Pharmaceuticals, Inc. TNF-α ribozymes
US5616488A (en) * 1992-12-07 1997-04-01 Ribozyme Pharmaceuticals, Inc. IL-5 targeted ribozymes
US5837542A (en) 1992-12-07 1998-11-17 Ribozyme Pharmaceuticals, Inc. Intercellular adhesion molecule-1 (ICAM-1) ribozymes
WO1995006731A2 (fr) 1993-09-02 1995-03-09 Ribozyme Pharmaceuticals, Inc. Acide nucleique enzymatique contenant un non-nucleotide
ES2127948T3 (es) * 1993-10-27 1999-05-01 Ribozyme Pharm Inc Oligonucleotidos modificados en la posicion 2'-amido y 2'-peptido.
AU687479B2 (en) 1993-11-08 1998-02-26 Ribozyme Pharmaceuticals, Inc. Base-modified enzymatic nucleic acid
DE69431564D1 (de) * 1993-11-12 2002-11-21 Ribozyme Pharm Inc Reagenz zur behandlung von arthritischen zustaenden
EP0746614A1 (fr) * 1994-02-23 1996-12-11 Ribozyme Pharmaceuticals, Inc. Procede et reactif inhibiteur de l'expression de genes concernant une affection
US5631359A (en) * 1994-10-11 1997-05-20 Ribozyme Pharmaceuticals, Inc. Hairpin ribozymes
US5902880A (en) * 1994-08-19 1999-05-11 Ribozyme Pharmaceuticals, Inc. RNA polymerase III-based expression of therapeutic RNAs
US5693532A (en) * 1994-11-04 1997-12-02 Ribozyme Pharmaceuticals, Inc. Respiratory syncytial virus ribozymes
US5639647A (en) * 1994-03-29 1997-06-17 Ribozyme Pharmaceuticals, Inc. 2'-deoxy-2'alkylnucleotide containing nucleic acid
US5834440A (en) * 1994-03-07 1998-11-10 Immusol Incorporated Ribozyme therapy for the inhibition of restenosis
US5627053A (en) * 1994-03-29 1997-05-06 Ribozyme Pharmaceuticals, Inc. 2'deoxy-2'-alkylnucleotide containing nucleic acid
US6103890A (en) * 1994-05-18 2000-08-15 Ribozyme Pharmaceuticals, Inc. Enzymatic nucleic acids that cleave C-fos
US20030026782A1 (en) * 1995-02-07 2003-02-06 Arthur M. Krieg Immunomodulatory oligonucleotides
US6350934B1 (en) 1994-09-02 2002-02-26 Ribozyme Pharmaceuticals, Inc. Nucleic acid encoding delta-9 desaturase
US5599706A (en) * 1994-09-23 1997-02-04 Stinchcomb; Dan T. Ribozymes targeted to apo(a) mRNA
US5672501A (en) * 1994-12-23 1997-09-30 Ribozyme Pharmaceuticals, Inc. Base-modified enzymatic nucleic acid
AU4412096A (en) * 1994-12-13 1996-07-03 Ribozyme Pharmaceuticals, Inc. Method and reagent for treatment of arthritic conditions, induction of graft tolerance and reversal of immune responses
US5716824A (en) 1995-04-20 1998-02-10 Ribozyme Pharmaceuticals, Inc. 2'-O-alkylthioalkyl and 2-C-alkylthioalkyl-containing enzymatic nucleic acids (ribozymes)
US5705388A (en) * 1994-12-23 1998-01-06 Ribozyme Pharmaceuticals, Inc. CETP Ribozymes
US5663064A (en) * 1995-01-13 1997-09-02 University Of Vermont Ribozymes with RNA protein binding site
JP3844501B2 (ja) 1995-02-08 2006-11-15 タカラバイオ株式会社 癌の制御
US5877021A (en) 1995-07-07 1999-03-02 Ribozyme Pharmaceuticals, Inc. B7-1 targeted ribozymes
US6346398B1 (en) 1995-10-26 2002-02-12 Ribozyme Pharmaceuticals, Inc. Method and reagent for the treatment of diseases or conditions related to levels of vascular endothelial growth factor receptor
US7034009B2 (en) 1995-10-26 2006-04-25 Sirna Therapeutics, Inc. Enzymatic nucleic acid-mediated treatment of ocular diseases or conditions related to levels of vascular endothelial growth factor receptor (VEGF-R)
US5998203A (en) 1996-04-16 1999-12-07 Ribozyme Pharmaceuticals, Inc. Enzymatic nucleic acids containing 5'-and/or 3'-cap structures
EP0914421A2 (fr) 1996-02-29 1999-05-12 Immusol, Inc. Ribozymes du virus de l'hepatite c
AU721758B2 (en) * 1996-04-02 2000-07-13 Gene Shears Pty. Limited Asymmetric hammerhead ribozymes
CN1254376A (zh) 1996-10-01 2000-05-24 杰龙公司 端粒酶逆转录酶
US5807743A (en) * 1996-12-03 1998-09-15 Ribozyme Pharmaceuticals, Inc. Interleukin-2 receptor gamma-chain ribozymes
US7008776B1 (en) 1996-12-06 2006-03-07 Aventis Pharmaceuticals Inc. Compositions and methods for effecting the levels of high density lipoprotein (HDL) cholesterol and apolipoprotein AI very low density lipoprotein (VLDL) cholesterol and low density lipoprotein (LDL) cholesterol
US6248878B1 (en) 1996-12-24 2001-06-19 Ribozyme Pharmaceuticals, Inc. Nucleoside analogs
US6057156A (en) 1997-01-31 2000-05-02 Robozyme Pharmaceuticals, Inc. Enzymatic nucleic acid treatment of diseases or conditions related to levels of epidermal growth factor receptors
US6159951A (en) * 1997-02-13 2000-12-12 Ribozyme Pharmaceuticals Inc. 2'-O-amino-containing nucleoside analogs and polynucleotides
AU7071598A (en) * 1997-04-10 1998-10-30 Erasmus University Rotterdam Diagnosis method and reagents
US5942395A (en) * 1997-05-09 1999-08-24 Universite De Montreal Hybrid ribozymes and methods of use
US6280936B1 (en) 1997-06-09 2001-08-28 Ribozyme Pharmaceuticals, Inc. Method for screening nucleic acid catalysts
US6548657B1 (en) 1997-06-09 2003-04-15 Ribozyme Pharmaceuticals, Inc. Method for screening nucleic acid catalysts
US6183959B1 (en) 1997-07-03 2001-02-06 Ribozyme Pharmaceuticals, Inc. Method for target site selection and discovery
US6316612B1 (en) 1997-08-22 2001-11-13 Ribozyme Pharmaceuticals, Inc. Xylofuranosly-containing nucleoside phosphoramidites and polynucleotides
JP2001518292A (ja) * 1997-09-22 2001-10-16 マックス−プランク−ゲゼルシャフト・ツア・フェルデルング・デア・ヴィッセンシャフテン・エー・ファオ エンドヌクレアーゼ活性を有する核酸触媒
US6656731B1 (en) 1997-09-22 2003-12-02 Max Planck Gesellschaft Zur Forderung Der Wissenschaften E.V. Nucleic acid catalysts with endonuclease activity
AU9509798A (en) * 1997-09-25 1999-04-12 University Of Florida Antisense oligonucleotide compositions targeted to angiotensi n converting enzy me mrna and methods of use
US6127535A (en) * 1997-11-05 2000-10-03 Ribozyme Pharmaceuticals, Inc. Nucleoside triphosphates and their incorporation into oligonucleotides
US6617438B1 (en) 1997-11-05 2003-09-09 Sirna Therapeutics, Inc. Oligoribonucleotides with enzymatic activity
PT1040192E (pt) 1997-12-18 2006-12-29 Monsanto Technology Llc Plantas transgénicas resistentes a insectos e métodos para melhoramento da actividade 8-endotoxina contra insectos
US20020147143A1 (en) 1998-03-18 2002-10-10 Corixa Corporation Compositions and methods for the therapy and diagnosis of lung cancer
US6200754B1 (en) 1998-03-19 2001-03-13 Variagenics, Inc. Inhibitors of alternative alleles of genes encoding products that mediate cell response to environmental changes
WO1999054459A2 (fr) 1998-04-20 1999-10-28 Ribozyme Pharmaceuticals, Inc. Molecules d'acides nucleiques presentant de nouvelles compositions chimiques capables de moduler l'expression genique
CA2230203A1 (fr) 1998-04-29 1999-10-29 Universite De Sherbrooke Ribozyme delta pour le clivage de l'arn
US6080580A (en) * 1998-10-05 2000-06-27 Isis Pharmaceuticals Inc. Antisense oligonucleotide modulation of tumor necrosis factor-α (TNF-α) expression
US6228642B1 (en) * 1998-10-05 2001-05-08 Isis Pharmaceuticals, Inc. Antisense oligonucleotide modulation of tumor necrosis factor-(α) (TNF-α) expression
US6995259B1 (en) 1998-10-23 2006-02-07 Sirna Therapeutics, Inc. Method for the chemical synthesis of oligonucleotides
WO2000066780A2 (fr) 1999-04-30 2000-11-09 University Of Florida Ribozymes fournis par virus adeno-associes pour la decouverte de fonctions geniques
WO2001032625A1 (fr) 1999-11-03 2001-05-10 Du Pont Pharmaceuticals Company Tetrahydroisoquinolines a substitution aryle et heteroaryle et utilisation de ces composes pour bloquer le recaptage de norepinephrine, de dopamine et de serotonine
US7163949B1 (en) 1999-11-03 2007-01-16 Amr Technology, Inc. 4-phenyl substituted tetrahydroisoquinolines and use thereof
US6831171B2 (en) 2000-02-08 2004-12-14 Yale University Nucleic acid catalysts with endonuclease activity
US8273866B2 (en) 2002-02-20 2012-09-25 Merck Sharp & Dohme Corp. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (SINA)
US20070026394A1 (en) 2000-02-11 2007-02-01 Lawrence Blatt Modulation of gene expression associated with inflammation proliferation and neurite outgrowth using nucleic acid based technologies
US8202979B2 (en) 2002-02-20 2012-06-19 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid
US20040002068A1 (en) 2000-03-01 2004-01-01 Corixa Corporation Compositions and methods for the detection, diagnosis and therapy of hematological malignancies
US7153839B2 (en) 2000-03-24 2006-12-26 Biocell Laboratories Method of protecting erythricytes, in particular for improvement of blood cytopenia
US6737271B1 (en) * 2000-03-24 2004-05-18 Biocell Laboratories Compound, composition and method for the treatment of inflammatory and inflammatory-related disorders
GB0018307D0 (en) 2000-07-26 2000-09-13 Aventis Pharm Prod Inc Polypeptides
NZ521786A (en) 2000-03-31 2004-07-30 Aventis Pharma Inc NFIF-14b and NFIF-7a (nuclear factor kappa B inducing factor) polypeptides and nulciec acids and their use in compositions for inhibiting inflammation
EP1319069B1 (fr) 2000-06-28 2008-05-21 Corixa Corporation Compositions et methodes pour le traitement et le diagnostic du cancer du poumon
JP5132864B2 (ja) 2000-07-11 2013-01-30 アルバニー モレキュラー リサーチ， インコーポレイテッド ４−フェニル置換テトラヒドロイソキノリンおよびその使用方法
US7754435B2 (en) 2000-09-21 2010-07-13 Philadelphia Health And Education Corporation Prostate cancer-related compositions, methods, and kits based on DNA macroarray proteomics platforms
WO2002072790A2 (fr) 2001-03-14 2002-09-19 Myriad Genetics, Inc Interaction tsg101-gag et son utilisation
US6613083B2 (en) 2001-05-02 2003-09-02 Eckhard Alt Stent device and method
CA2446788A1 (fr) 2001-05-09 2002-11-14 Corixa Corporation Compositions et procedes pour la therapie et le diagnostic du cancer de la prostate
US9994853B2 (en) 2001-05-18 2018-06-12 Sirna Therapeutics, Inc. Chemically modified multifunctional short interfering nucleic acid molecules that mediate RNA interference
US20050148530A1 (en) 2002-02-20 2005-07-07 Sirna Therapeutics, Inc. RNA interference mediated inhibition of vascular endothelial growth factor and vascular endothelial growth factor receptor gene expression using short interfering nucleic acid (siNA)
US7517864B2 (en) 2001-05-18 2009-04-14 Sirna Therapeutics, Inc. RNA interference mediated inhibition of vascular endothelial growth factor and vascular endothelial growth factor receptor gene expression using short interfering nucleic acid (siNA)
US7205399B1 (en) 2001-07-06 2007-04-17 Sirna Therapeutics, Inc. Methods and reagents for oligonucleotide synthesis
EP1581119B1 (fr) 2001-12-17 2013-01-30 Corixa Corporation Compositions et procedes applicables a la therapie et au diagnostic pour la maladie intestinale inflammatoire
AU2003207708A1 (en) 2002-02-20 2003-09-09 Sirna Therapeutics, Inc. Rna interference mediated inhibition of map kinase genes
US9657294B2 (en) 2002-02-20 2017-05-23 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)
US9181551B2 (en) 2002-02-20 2015-11-10 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)
US7923547B2 (en) 2002-09-05 2011-04-12 Sirna Therapeutics, Inc. RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)
US20080260744A1 (en) 2002-09-09 2008-10-23 Omeros Corporation G protein coupled receptors and uses thereof
JP2006507841A (ja) * 2002-11-14 2006-03-09 ダーマコン， インコーポレイテッド 機能的ｓｉＲＮＡおよび超機能的ｓｉＲＮＡ
US7829694B2 (en) 2002-11-26 2010-11-09 Medtronic, Inc. Treatment of neurodegenerative disease through intracranial delivery of siRNA
US7605249B2 (en) 2002-11-26 2009-10-20 Medtronic, Inc. Treatment of neurodegenerative disease through intracranial delivery of siRNA
US7618948B2 (en) 2002-11-26 2009-11-17 Medtronic, Inc. Devices, systems and methods for improving and/or cognitive function through brain delivery of siRNA
US7994149B2 (en) 2003-02-03 2011-08-09 Medtronic, Inc. Method for treatment of Huntington's disease through intracranial delivery of sirna
US7700280B2 (en) 2003-12-31 2010-04-20 The Penn State Research Foundation Methods for assessing cisplatin resistance, disease progression, and treatment efficacy in ovarian cancer
ES2423060T3 (es) 2004-03-12 2013-09-17 Alnylam Pharmaceuticals, Inc. Agentes iRNA que tienen como diana al VEGF
SI1745295T1 (sl) 2004-04-20 2011-01-31 Galapagos Nv Metode, sestavljanje in preizkuĺ anje spojine za zaviranje tvorbe proteina amiloid-beta
CA2562833A1 (fr) 2004-04-27 2005-11-03 Galapagos N.V. Procedes, agents, et analyses de criblage de composes permettant d'induire une differenciation de cellules mammaliennes non differenciees en osteoblastes
US20060040882A1 (en) 2004-05-04 2006-02-23 Lishan Chen Compostions and methods for enhancing delivery of nucleic acids into cells and for modifying expression of target genes in cells
US10508277B2 (en) 2004-05-24 2019-12-17 Sirna Therapeutics, Inc. Chemically modified multifunctional short interfering nucleic acid molecules that mediate RNA interference
AU2005248167B2 (en) 2004-05-26 2010-11-25 Anima Cell Metrology Methods for evaluating ribonucleotide sequences
WO2005121778A2 (fr) 2004-06-14 2005-12-22 Galapagos N.V. Procedes d'identification, et composes utiles pour le traitement de maladies degeneratives et inflammatoires
JP2008503712A (ja) 2004-06-21 2008-02-07 ガラパゴス・ナムローゼ・フェンノートシャップ 骨関節炎治療の方法及び手段
EP2298896A1 (fr) 2004-06-22 2011-03-23 The Board of Trustees of the University of Illinois Procédés pour inhiber la prolifération des cellules tumorales avec FOXM1 ARNsi
USRE48960E1 (en) 2004-06-28 2022-03-08 The University Of Western Australia Antisense oligonucleotides for inducing exon skipping and methods of use thereof
PL1766010T3 (pl) 2004-06-28 2011-07-29 Univ Western Australia Antysensowne oligonukleotydy do indukcji pominięcia egzonu i sposoby ich zastosowania
AU2005274927B2 (en) 2004-07-15 2011-11-03 Albany Molecular Research, Inc. Aryl-and heteroaryl-substituted tetrahydroisoquinolines and use thereof to block reuptake of norepinephrine, dopamine, and serotonin
US7485468B2 (en) 2004-10-15 2009-02-03 Galapagos Bv Molecular targets and compounds, and methods to identify the same, useful in the treatment of joint degenerative and inflammatory diseases
ATE541928T1 (de) 2005-03-31 2012-02-15 Calando Pharmaceuticals Inc Inhibitoren der untereinheit 2 der ribonukleotidreduktase und ihre verwendungen
US8362075B2 (en) 2005-05-17 2013-01-29 Merck Sharp & Dohme Corp. Cyclohexyl sulphones for treatment of cancer
EP1896084A4 (fr) 2005-06-27 2010-10-20 Alnylam Pharmaceuticals Inc Modulation de l'arni de hif-1 et ses applications thérapeutiques
US9133517B2 (en) 2005-06-28 2015-09-15 Medtronics, Inc. Methods and sequences to preferentially suppress expression of mutated huntingtin
KR101594898B1 (ko) 2005-07-15 2016-02-18 알바니 몰레큘라 리써치, 인크. 아릴- 및 헤테로아릴-치환된 테트라히드로벤자제핀, 및 노르에피네프린, 도파민 및 세로토닌의 재흡수를 차단하기 위한 용도
US20070213292A1 (en) 2005-08-10 2007-09-13 The Rockefeller University Chemically modified oligonucleotides for use in modulating micro RNA and uses thereof
KR20080048502A (ko) 2005-09-29 2008-06-02 머크 앤드 캄파니 인코포레이티드 멜라노코르틴-4 수용체 조절제로서의 아실화스피로피페리딘 유도체
EP2671954B1 (fr) 2006-01-20 2018-05-16 Cell Signaling Technology, Inc. Translocation et kinase ROS mutante dans un cancer du poumon non à petites cellules chez un être humain
US20120208824A1 (en) 2006-01-20 2012-08-16 Cell Signaling Technology, Inc. ROS Kinase in Lung Cancer
GB0603041D0 (en) 2006-02-15 2006-03-29 Angeletti P Ist Richerche Bio Therapeutic compounds
WO2007109097A2 (fr) 2006-03-16 2007-09-27 Alnylam Pharmaceuticals, Inc. modulation ARNi des TGF-beta et usages therapeutiques
CA2647728C (fr) 2006-03-31 2014-12-09 Alnylam Pharmaceuticals, Inc. Compositions et methodes destinees a inhiber l'expression du gene eg5
US7741469B2 (en) 2006-04-05 2010-06-22 University Of Iowa Research Foundation Compositions for treating hearing loss and methods of use thereof
CA2648718A1 (fr) 2006-04-07 2007-10-18 The Research Foundation Of State University Of New York Polypeptides du recepteur de la transcobalamine, acides nucleiques et modulateurs associes, procedes d'utilisation associes destines a moduler la croissance cellulaire et a traiter le cancer et la deficience en cobalamine
EP2447359B1 (fr) 2006-04-14 2015-11-04 Cell Signaling Technology, Inc. Défauts de gène et ALK kinase mutante dans des tumeurs solides humaines
US8377448B2 (en) 2006-05-15 2013-02-19 The Board Of Trustees Of The Leland Standford Junior University CD47 related compositions and methods for treating immunological diseases and disorders
US9273356B2 (en) 2006-05-24 2016-03-01 Medtronic, Inc. Methods and kits for linking polymorphic sequences to expanded repeat mutations
US8598333B2 (en) 2006-05-26 2013-12-03 Alnylam Pharmaceuticals, Inc. SiRNA silencing of genes expressed in cancer
WO2008054561A2 (fr) 2006-07-11 2008-05-08 University Of Medicine And Dentistry Of New Jersey Protéines, acides nucléiques encodant celles-ci et leurs procédés d'utilisation associés
RU2553561C2 (ru) 2006-07-21 2015-06-20 Сайленс Терапьютикс Аг Средства ингибирования экспрессии протеинкиназы 3
EP1900749A1 (fr) 2006-09-12 2008-03-19 Institut National De La Sante Et De La Recherche Medicale (Inserm) Acide nucléique pour l'expression d'un polynucléotide d'interêt dans des cellules cancéreuses de mammiferès
US8173629B2 (en) 2006-09-22 2012-05-08 Merck Sharp & Dohme Corp. Method of treatment using fatty acid synthesis inhibitors
AU2007302803A1 (en) 2006-10-02 2008-04-10 Aprea Ab RNA interference against Wrap53 to treat hyperproliferative diseases
CN105061580B (zh) 2006-10-03 2020-06-05 罗格斯新泽西州立大学 多肽和药物组合物及其应用
US20110218176A1 (en) 2006-11-01 2011-09-08 Barbara Brooke Jennings-Spring Compounds, methods, and treatments for abnormal signaling pathways for prenatal and postnatal development
US9375440B2 (en) 2006-11-03 2016-06-28 Medtronic, Inc. Compositions and methods for making therapies delivered by viral vectors reversible for safety and allele-specificity
WO2009024834A2 (fr) 2006-12-05 2009-02-26 Rosetta Genomics Ltd Acides nucléiques impliques dans les infections virales
EA016079B1 (ru) 2007-01-10 2012-01-30 Институто Ди Ричерке Ди Биолоджиа Молеколаре П. Анджелетти Спа Амидзамещенные индазолы в качестве ингибиторов поли(adp-рибоза)полимеразы (parp)
WO2008092099A2 (fr) 2007-01-26 2008-07-31 Rosetta Genomics, Ltd. Compositions et procédés de traitement de malignités hématopoïétiques
ATE553196T1 (de) 2007-02-27 2012-04-15 Rosetta Genomics Ltd Zusammensetzung und verfahren zur modulation von zellproliferation und zelltod
US9006206B2 (en) 2007-02-27 2015-04-14 Rosetta Genomics Ltd. Composition and methods for modulating cell proliferation and cell death
WO2008106551A2 (fr) 2007-02-28 2008-09-04 The Govt. Of The U.S.A. As Represented By The Secretary Of The Dept. Of Health & Human Serv. Polypeptides brachyury et procédés d'utilisation.
WO2008109377A1 (fr) 2007-03-02 2008-09-12 Mdrna, Inc. Composés d'acide nucléique permettant d'inhiber l'expression de gène de la famille vegf et utilisations de ceux-ci
JP5319518B2 (ja) 2007-04-02 2013-10-16 Ｍｓｄ株式会社 インドールジオン誘導体
ES2529790T3 (es) 2007-04-13 2015-02-25 Dana-Farber Cancer Institute, Inc. Métodos de tratamiento de cáncer resistente a agentes terapéuticos de ERBB
LT2494993T (lt) 2007-05-04 2018-12-27 Marina Biotech, Inc. Aminorūgščių lipidai ir jų panaudojimas
HRP20140405T1 (hr) 2007-06-20 2014-06-20 Galapagos N.V. Molekularni ciljevi i metode za identificiranje spojeva za lijeäśenje degenerativnih bolesti kostiju i zglobova
AU2008269154B2 (en) 2007-06-27 2014-06-12 Merck Sharp & Dohme Llc 4-carboxybenzylamino derivatives as histone deacetylase inhibitors
AU2008309146B2 (en) 2007-10-09 2014-06-12 Anima Cell Metrology, Inc. Systems and methods for measuring translation activity in viable cells
EP3741851A1 (fr) 2007-10-18 2020-11-25 Cell Signaling Technology, Inc. Translocation et kinase ros mutante dans un carcinome pulmonaire non à petites cellules humaines
CA2700215A1 (fr) 2007-10-31 2009-05-07 Rosetta Genomics Ltd. Diagnostic et pronostic de cancers specifiques
EP2217716A4 (fr) 2007-11-09 2011-02-09 Salk Inst For Biological Studi Utilisation d'inhibiteurs de récepteurs tam en tant qu'antimicrobiens
CN102006890A (zh) 2007-12-04 2011-04-06 阿尔尼拉姆医药品有限公司 靶向脂质
JP5749494B2 (ja) 2008-01-02 2015-07-15 テクミラ ファーマシューティカルズ コーポレイション 核酸の送達のための改善された組成物および方法
CN102014631A (zh) 2008-03-03 2011-04-13 泰格尔医药科技公司 酪氨酸激酶抑制剂
WO2009123764A2 (fr) 2008-04-04 2009-10-08 Calando Pharmaceuticals, Inc. Compositions et utilisation d'inhibiteurs d'epas1
EP3604533A1 (fr) 2008-04-11 2020-02-05 Arbutus Biopharma Corporation Délivrance spécifique à un site d'acides nucléiques en combinant des ligands de ciblage avec des composants endosomolytiques
NZ588583A (en) 2008-04-15 2012-08-31 Protiva Biotherapeutics Inc Novel lipid formulations for nucleic acid delivery
WO2009134443A2 (fr) 2008-05-02 2009-11-05 The Brigham And Women's Hospital, Inc. Apoptose et silençage traductionnel à induction par l'arn
AU2009246134B2 (en) 2008-05-16 2016-03-03 The Children's Hospital Of Philadelphia Genetic alterations on chromosomes 21q, 6q and 15q and methods of use thereof for the diagnosis and treatment of type I diabetes
JP2011521649A (ja) 2008-05-30 2011-07-28 イェール ユニバーシティー 遺伝子発現を改変するための標的化オリゴヌクレオチド組成物
US8222221B2 (en) 2008-06-04 2012-07-17 The Board Of Regents Of The University Of Texas System Modulation of gene expression through endogenous small RNA targeting of gene promoters
US9156812B2 (en) 2008-06-04 2015-10-13 Bristol-Myers Squibb Company Crystalline form of 6-[(4S)-2-methyl-4-(2-naphthyl)-1,2,3,4-tetrahydroisoquinolin-7-yl]pyridazin-3-amine
WO2010029118A1 (fr) 2008-09-11 2010-03-18 Biofocus Dpi B.V. Procédé d'identification de composés utiles pour accroître l'activité fonctionnelle et l'expression à la surface cellulaire d'un régulateur de la conductance transmembranaire de la mucoviscidose mutant associé la mucoviscidose (mv)
EP2342616A2 (fr) 2008-09-23 2011-07-13 Alnylam Pharmaceuticals Inc. Modifications chimiques de monomères et d oligonucléotides par cycloaddition
JP5777519B2 (ja) 2008-10-09 2015-09-09 テクミラ ファーマシューティカルズ コーポレイション 改良されたアミノ脂質および核酸の送達方法
EP2349210B1 (fr) 2008-10-16 2015-03-18 Marina Biotech, Inc. Procédés et compositions pour une administration liposomale et efficace de produits thérapeutiques de silençage de gène
CN105779453A (zh) 2008-10-24 2016-07-20 萨雷普塔治疗公司 用于dmd的多外显子跳跃组合物
IT1391866B1 (it) * 2008-10-30 2012-01-27 Mastelli S R L Composizione iniettabile di polinucleotidi per il trattamento di patologie osteoarticolari.
EP4241767B1 (fr) 2008-11-10 2024-10-23 Arbutus Biopharma Corporation Nouveaux lipides et compositions d'administration d'agents thérapeutiques
US20120101148A1 (en) 2009-01-29 2012-04-26 Alnylam Pharmaceuticals, Inc. lipid formulation
JP5979877B2 (ja) 2009-02-12 2016-08-31 セル・シグナリング・テクノロジー・インコーポレイテツド ヒト癌における変異ｒｏｓ発現
WO2010091878A2 (fr) 2009-02-13 2010-08-19 Silence Therapeutics Ag Moyens pour inhiber l'expression du opa1
JP2012517815A (ja) 2009-02-18 2012-08-09 サイレンス・セラピューティクス・アーゲー Ａｎｇ２の発現を阻害するための手段
EP2398480A1 (fr) 2009-02-19 2011-12-28 Galapagos N.V. Procédé pour identifier des composés utiles pour diagnostiquer et traiter des maladies impliquant une inflammation
JP2012517822A (ja) 2009-02-19 2012-08-09 ガラパゴス・ナムローゼ・フェンノートシャップ 炎症を包含する疾患の診断及び治療に有用な同定方法及び化合物
JP2012517821A (ja) 2009-02-19 2012-08-09 ガラパゴス・ナムローゼ・フェンノートシャップ 炎症を包含する疾患の診断及び治療に有用な同定方法及び化合物
EP3424939A1 (fr) 2009-03-02 2019-01-09 Alnylam Pharmaceuticals Inc. Modifications chimiques d'acide nucléique
US10702583B2 (en) 2009-03-11 2020-07-07 Promedior, Inc. Treatment methods for autoimmune disorders
JP5797565B2 (ja) 2009-03-11 2015-10-21 プロメディオール， インコーポレイテッド 過敏性障害（ｈｙｐｅｒｓｅｎｓｉｔｉｖｅ ｄｉｓｏｒｄｅｒ）に対する処置および診断方法
CA2755773A1 (fr) 2009-03-19 2010-09-23 Merck Sharp & Dohme Corp. Inhibition mediee par arn interference de l'expression genique de btb et de l'homologie cnc 1, facteur de transcription 1 de fermeture eclair de leucine basique (bach 1), utilisant une liste de sequences d'acide nucleique interferant court (ansi)
WO2010107957A2 (fr) 2009-03-19 2010-09-23 Merck Sharp & Dohme Corp. Inhibition induite par arn interférence d'une expression génique (gata3) d'une protéine de liaison gata au moyen d'un acide nucléique interférent court
JP2012520683A (ja) 2009-03-19 2012-09-10 メルク・シャープ・エンド・ドーム・コーポレイション 低分子干渉核酸（ｓｉＮＡ）を用いた結合組織増殖因子（ＣＴＧＦ）遺伝子発現のＲＮＡ干渉媒介性阻害
WO2010107958A1 (fr) 2009-03-19 2010-09-23 Merck Sharp & Dohme Corp. INHIBITION INDUITE PAR ARN INTERFÉRENCE DE L'EXPRESSION DU GÈNE TRANSDUCTEUR DE SIGNAL ET ACTIVITATEUR DE TRANSCRIPTION 6 (STAT6) AU MOYEN D'UN ACIDE NUCLÉIQUE INTERFÉRENT COURT (ANsi)
WO2010111471A2 (fr) 2009-03-27 2010-09-30 Merck Sharp & Dohme Corp. Inhibition par interférence arn de l'expression du gène du signal transducteur et activateur de la transcription 1 (stat1) au moyen d'un acide nucléique interférent court (ansi)
WO2010111468A2 (fr) 2009-03-27 2010-09-30 Merck Sharp & Dohme Corp. INHIBITION PAR INTERFÉRENCE ARN DE L'EXPRESSION DU GÈNE DE LA CHAÎNE BÊTA DU FACTEUR DE CROISSANCE DES NERFS (NGFß) AU MOYEN D'UN ACIDE NUCLÉIQUE INTERFÉRENT COURT (ANSI)
EP2411520A2 (fr) 2009-03-27 2012-02-01 Merck Sharp&Dohme Corp. Inhibition à médiation par l'interférence arn de l'expression du gène de la lymphopoïétine stromale thymique (tslp) faisant appel à de courts acides nucléiques interférents (ansi)
MX2011010072A (es) 2009-03-27 2011-10-06 Merck Sharp & Dohme Inhibicion mediada por interferencia de acido ribonucleico de la expresion del gen de la molecula de adhesion intercelular 1 usando acido nucleico corto de interferencia.
US20120010272A1 (en) 2009-03-27 2012-01-12 Merck Sharp & Dohme Corp. RNA Interference Mediated Inhibition of Apoptosis Signal-Regulating Kinase 1 (ASK1) Gene Expression Using Short Interfering Nucleic Acid (siNA)
WO2010112569A1 (fr) 2009-03-31 2010-10-07 Robert Zimmermann Modulation de lipase de triglycérides d'adipose pour la prévention et le traitement d'une cachexie, d'une perte de poids et d'une atrophie musculaire et procédés de criblage s'y rapportant
US8993509B2 (en) 2009-03-31 2015-03-31 Robert Zimmerman Method for treatment of cachexia by administering inhibitors of adipose triglyceride lipase expression or activity
JP2012522977A (ja) 2009-04-01 2012-09-27 ガラパゴス・ナムローゼ・フェンノートシャップ 変形性関節症の治療のための方法及び手段
WO2010114780A1 (fr) 2009-04-01 2010-10-07 Merck Sharp & Dohme Corp. Inhibiteurs de l'activité akt
US8815586B2 (en) 2009-04-24 2014-08-26 The Board Of Regents Of The University Of Texas System Modulation of gene expression using oligomers that target gene regions downstream of 3′ untranslated regions
SG176553A1 (en) 2009-05-05 2012-01-30 Muthiah Manoharan Lipid compositions
CA2760706C (fr) 2009-05-05 2019-08-06 Alnylam Pharmaceuticals, Inc. Procedes d'administration d'oligonucleotides en direction de cellules immunitaires
US8802696B2 (en) 2009-05-12 2014-08-12 Albany Molecular Research, Inc. 7-([1,2,4]triazolo[1,5-a]pyridin-6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoqu inoli and use thereof
WO2010132487A1 (fr) 2009-05-12 2010-11-18 Bristol-Myers Squibb Company Formes cristallines de (s)-7-([1,2,4]triazolo[1,5-a] pyridin -6-yl)-4-(3,4-dichlorophenyl)-1,2,3,4-tetrahydroisoquinoline et leurs utilisations
CN102458123A (zh) 2009-05-12 2012-05-16 阿尔巴尼分子研究公司 芳基、杂芳基和杂环取代的四氢异喹啉及其用途
UA110323C2 (en) 2009-06-04 2015-12-25 Promedior Inc Derivative of serum amyloid p and their receipt and application
TR201811076T4 (tr) 2009-06-10 2018-08-27 Arbutus Biopharma Corp Geliştirilmiş lipit formulasyonu.
US8283333B2 (en) 2009-07-01 2012-10-09 Protiva Biotherapeutics, Inc. Lipid formulations for nucleic acid delivery
WO2011005861A1 (fr) 2009-07-07 2011-01-13 Alnylam Pharmaceuticals, Inc. Coiffes d’extrémité d’oligonucléotides
WO2011015572A1 (fr) 2009-08-03 2011-02-10 Galapagos Nv Cibles et composés moléculaires, et procédés pour leur identification utiles dans le traitement de maladies neurodégénératives
WO2011015573A1 (fr) 2009-08-03 2011-02-10 Galapagos Nv Cibles et composés moléculaires, et procédés pour les identifier, utiles dans le traitement de maladies neurodégénératives
AU2010299444B2 (en) 2009-09-24 2015-12-10 Anima Cell Metrology, Inc. Systems and methods for measuring translation of target proteins in cells
JP5099731B1 (ja) 2009-10-14 2012-12-19 メルク・シャープ・アンド・ドーム・コーポレーション ｐ５３活性を増大する置換ピペリジン及びその使用
US20120270930A1 (en) 2009-10-29 2012-10-25 Academisch Ziekenhuis Leiden H.O.D.N. Lumc Methods and compositions for dysferlin exon-skipping
BR122023023194A2 (pt) 2009-11-12 2023-12-26 The University Of Western Australia Oligonucleotídeo antissentido, composição e respectivo uso
JP5991922B2 (ja) 2009-11-13 2016-09-14 サレプタ セラピューティクス， インコーポレイテッド アンチセンス抗ウイルス性化合物およびインフルエンザウイルス感染を処置するための方法
WO2011071860A2 (fr) 2009-12-07 2011-06-16 Alnylam Pharmaceuticals, Inc. Compositions utilisées pour l'administration d'acides nucléiques
JP5685267B2 (ja) 2009-12-09 2015-03-18 日東電工株式会社 Ｈｓｐ４７発現の調節
EP3494963A1 (fr) 2009-12-18 2019-06-12 The University of British Columbia Procédés et compositions d'administration d'acides nucléiques
WO2011094580A2 (fr) 2010-01-28 2011-08-04 Alnylam Pharmaceuticals, Inc. Cuivre chélaté à utiliser dans la préparation d'oligonucléotides conjugués
EP2545173A2 (fr) 2010-03-12 2013-01-16 Sarepta Therapeutics, Inc. Modulation anti-sens de récepteurs hormonaux nucléaires
WO2011120023A1 (fr) 2010-03-26 2011-09-29 Marina Biotech, Inc. Composés d'acide nucléique pour inhiber l'expression de gène de survivine et utilisations de ceux-ci
WO2011123621A2 (fr) 2010-04-01 2011-10-06 Alnylam Pharmaceuticals Inc. Monomères modifiés en 2' et 5' et oligonucléotides
AU2011237630B2 (en) 2010-04-06 2016-01-21 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of CD274/PD-L1 gene
WO2011133584A2 (fr) 2010-04-19 2011-10-27 Marina Biotech, Inc. Composés de type acide nucléique pour inhiber l'expression du gène hras et leurs utilisations
US20130260460A1 (en) 2010-04-22 2013-10-03 Isis Pharmaceuticals Inc Conformationally restricted dinucleotide monomers and oligonucleotides
WO2011133871A2 (fr) 2010-04-22 2011-10-27 Alnylam Pharmaceuticals, Inc. Dérivés d'extrémité 5'
US10913767B2 (en) 2010-04-22 2021-02-09 Alnylam Pharmaceuticals, Inc. Oligonucleotides comprising acyclic and abasic nucleosides and analogs
KR20200047642A (ko) 2010-04-23 2020-05-07 유니버시티 오브 플로리다 리서치 파운데이션, 인크. 레베르 선천성 흑내장-1(lca1)의 치료를 위한 재조합 아데노-관련된 바이러스-구아닐레이트 사이클라제 조성물 및 방법
WO2011139842A2 (fr) 2010-04-28 2011-11-10 Marina Biotech, Inc. Composés d'acide nucléique destinés à inhiber l'expression du gène fgfr3 et utilisations associées
ES2816898T3 (es) 2010-05-13 2021-04-06 Sarepta Therapeutics Inc Compuestos que modulan la actividad de señalización de las interleucinas 17 y 23
CA2800065A1 (fr) 2010-05-21 2011-11-24 Peptimed, Inc. Reactifs et methodes pour le traitement du cancer
US9944671B2 (en) 2010-06-02 2018-04-17 Alnylam Pharmaceuticals, Inc. Compositions and methods directed to treating liver fibrosis
US20130236968A1 (en) 2010-06-21 2013-09-12 Alnylam Pharmaceuticals, Inc. Multifunctional copolymers for nucleic acid delivery
RU2012153703A (ru) 2010-06-24 2014-07-27 Кварк Фармасьютикалс, Инк. Соединения двухцепочечных рнк к rhoa и их применение
EP2584903B1 (fr) 2010-06-24 2018-10-24 Merck Sharp & Dohme Corp. Nouveaux composés hétérocycliques utilisés comme inhibiteurs de erk
WO2012011110A2 (fr) 2010-07-22 2012-01-26 Ramot At Tel Aviv University Ltd. Systèmes et méthodes de détection du stress cellulaire
US20130323269A1 (en) 2010-07-30 2013-12-05 Muthiah Manoharan Methods and compositions for delivery of active agents
WO2012016188A2 (fr) 2010-07-30 2012-02-02 Alnylam Pharmaceuticals, Inc. Procédés et compositions pour l'administration d'agents actifs
AU2011285909B2 (en) 2010-08-02 2016-11-10 Sirna Therapeutics, Inc. RNA interference mediated inhibition of catenin (cadherin-associated protein), beta 1 (CTNNB1) gene expression using short interfering nucleic acid (siNA)
EP2601212A1 (fr) 2010-08-04 2013-06-12 Cizzle Biotechnology Limited Procédés et composés pour le diagnostic et le traitement du cancer
US20120101108A1 (en) 2010-08-06 2012-04-26 Cell Signaling Technology, Inc. Anaplastic Lymphoma Kinase In Kidney Cancer
EP2606134B1 (fr) 2010-08-17 2019-04-10 Sirna Therapeutics, Inc. Inhibition médiée par des arn interférents de l'expression génique du virus de l'hépatite b (vhb) à l'aide de petits acides nucléiques interférents (pani)
US8883801B2 (en) 2010-08-23 2014-11-11 Merck Sharp & Dohme Corp. Substituted pyrazolo[1,5-a]pyrimidines as mTOR inhibitors
WO2012027206A1 (fr) 2010-08-24 2012-03-01 Merck Sharp & Dohme Corp. Agents à base d'arni à un seul brin contenant une séquence intercalaire interne ne correspondant pas à un acide nucléique
US9233997B2 (en) 2010-08-26 2016-01-12 Sirna Therapeutics, Inc. RNA interference mediated inhibition of prolyl hydroxylase domain 2 (PHD2) gene expression using short interfering nucleic acid (siNA)
EP2613782B1 (fr) 2010-09-01 2016-11-02 Merck Sharp & Dohme Corp. Dérivés d'indazole utilisables en tant qu'inhibiteurs de la voie erk
US9242981B2 (en) 2010-09-16 2016-01-26 Merck Sharp & Dohme Corp. Fused pyrazole derivatives as novel ERK inhibitors
WO2012047631A2 (fr) 2010-09-27 2012-04-12 The Children's Hospital Of Philadelphia Compositions et procédés utiles dans le traitement et le diagnostic d'une maladie intestinale inflammatoire
SG10201508118WA (en) 2010-09-30 2015-11-27 Agency Science Tech & Res Methods and reagents for detection and treatment of esophageal metaplasia
US20140134231A1 (en) 2010-10-11 2014-05-15 Sanford-Burnham Medical Research Institute Mir-211 expression and related pathways in human melanoma
EP3766975A1 (fr) 2010-10-29 2021-01-20 Sirna Therapeutics, Inc. Inhibition au moyen d'interférence arn d'une expression de gène utilisant des acides nucléiques à petit interférent (sina)
EP2635308A2 (fr) 2010-11-01 2013-09-11 Peptimed, Inc. Compositions d'un système à base de peptides pour le ciblage spécifique de cellules
US9193973B2 (en) 2010-12-10 2015-11-24 Alynylam Pharmaceuticals, Inc. Compositions and methods for increasing erythropoietin (EPO) production
EP2648763A4 (fr) 2010-12-10 2014-05-14 Alnylam Pharmaceuticals Inc Compositions et procédés d'inhibition de l'expression des gènes klf-1 et bcl11a
EP2654748B1 (fr) 2010-12-21 2016-07-27 Merck Sharp & Dohme Corp. Dérivés d'indazole utiles en tant qu'inhibiteurs de erk
WO2012093384A1 (fr) 2011-01-03 2012-07-12 Rosetta Genomics Ltd. Compositions et procédés de traitement du cancer de l'ovaire
JP5902197B2 (ja) 2011-01-11 2016-04-13 アルニラム・ファーマシューティカルズ・インコーポレーテッド Ｐｅｇ化脂質および薬剤送達のためのそれらの使用
US20120263754A1 (en) 2011-02-15 2012-10-18 Immune Design Corp. Methods for Enhancing Immunogen Specific Immune Responses by Vectored Vaccines
EP2681314B1 (fr) 2011-03-03 2017-11-01 Quark Pharmaceuticals, Inc. Compositions et procédés pour traiter des maladies et des lésions pulmonaires
US8420617B2 (en) 2011-03-11 2013-04-16 Biocell Laboratories Multiantivirus compound, composition and method for treatment of virus diseases
MX360349B (es) 2011-03-29 2018-10-30 Alnylam Pharmaceuticals Inc Composiciones y metodos para inhibir la expresion del gen de proteasa transmembrana, serina 6 (tmprss6).
AU2012243039B2 (en) 2011-04-08 2017-07-13 Immune Design Corp. Immunogenic compositions and methods of using the compositions for inducing humoral and cellular immune responses
EP2699567A1 (fr) 2011-04-21 2014-02-26 Merck Sharp & Dohme Corp. Inhibiteurs du récepteur du facteur de croissance 1 analogue à l'insuline
TWI658830B (zh) 2011-06-08 2019-05-11 日東電工股份有限公司 Ｈｓｐ47表現調控強化用類視色素脂質體
US10196637B2 (en) 2011-06-08 2019-02-05 Nitto Denko Corporation Retinoid-lipid drug carrier
EP2723865B1 (fr) 2011-06-21 2019-03-27 Alnylam Pharmaceuticals, Inc. PROCÉDÉS DE DÉTERMINATION DE L'ACTIVITÉ DU RNAi CHEZ UN SUJET
EP3444348A1 (fr) 2011-06-21 2019-02-20 Alnylam Pharmaceuticals, Inc. Compositions d'arni faisant intervenir la protéine 3 de type angiopoïétine (angptl3) et leurs procédés d'utilisation
CN103608030A (zh) 2011-06-21 2014-02-26 昂科发克特公司 用于治疗和诊断癌症的组合物和方法
WO2012178033A2 (fr) 2011-06-23 2012-12-27 Alnylam Pharmaceuticals, Inc. Arnsi de serpina1 : compositions et méthodes de traitement
WO2013003697A1 (fr) 2011-06-30 2013-01-03 Trustees Of Boston University Méthode de régulation de croissance tumorale, d'angiogenèse et de métastase à l'aide du récepteur 1 riche en proline et contenant une immunoglobuline (igpr-1)
EP2732035A2 (fr) 2011-07-15 2014-05-21 Sarepta Therapeutics, Inc. Procédés et compositions pour manipulation de la traduction d'isoformes de protéines à partir de sites initiateurs alternatifs d'initiation
EP2557089A2 (fr) 2011-07-15 2013-02-13 Fundació Institut d'Investigació Biomèdica de Bellvitge (IDIBELL) Compositions et procédés d'immunomodulation
WO2013019857A2 (fr) 2011-08-01 2013-02-07 Alnylam Pharmaceuticals, Inc. Procédé permettant d'améliorer le taux de succès des greffes de cellules souches hématopoïétiques
EP2760477B1 (fr) 2011-09-27 2018-08-08 Alnylam Pharmaceuticals, Inc. Lipides di-aliphatiques pegylés substitués
WO2013063214A1 (fr) 2011-10-27 2013-05-02 Merck Sharp & Dohme Corp. Nouveaux composés qui sont des inhibiteurs d'erk
US10208086B2 (en) 2011-11-11 2019-02-19 Fred Hutchinson Cancer Research Center Cyclin A1-targeted T-cell immunotherapy for cancer
WO2013086441A2 (fr) 2011-12-08 2013-06-13 Sarepta Therapeutics, Inc. Analogues d'oligonucléotide ciblant lmna humain
WO2013090457A2 (fr) 2011-12-12 2013-06-20 Oncoimmunin Inc. Administration in vivo d'oligonucléotides
JP2016528161A (ja) 2012-01-12 2016-09-15 クォーク ファーマシューティカルズ インコーポレーティッドＱｕａｒｋ Ｐｈａｒｍａｃｅｕｔｉｃａｌｓ，Ｉｎｃ． 聴覚およびバランス障害を治療するための併用療法
EP2617434A1 (fr) 2012-01-20 2013-07-24 Laboratorios Del. Dr. Esteve, S.A. Immunogènes déficients en intégrase du VIH-1 et procédé pour le chargement de cellules dendritiques avec de tels immunogènes
WO2013123996A1 (fr) 2012-02-24 2013-08-29 Astrazeneca Uk Limited Nouveaux inhibiteurs d'arnsi de icam-1 humain
DK2817287T3 (da) 2012-02-24 2019-01-02 Arbutus Biopharma Corp Trialkyl kationisk lipid og metoder til anvendelse deraf
US9133461B2 (en) 2012-04-10 2015-09-15 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of the ALAS1 gene
ES2655842T3 (es) 2012-04-18 2018-02-21 Cell Signaling Technology, Inc. EGFR y ROS1 en el cáncer
US9127274B2 (en) 2012-04-26 2015-09-08 Alnylam Pharmaceuticals, Inc. Serpinc1 iRNA compositions and methods of use thereof
EP3358013B1 (fr) 2012-05-02 2020-06-24 Sirna Therapeutics, Inc. Compositions d'acide nucléique interférent court (sina)
CN112007045B (zh) 2012-07-13 2026-01-06 波涛生命科学有限公司 手性控制
WO2014018375A1 (fr) 2012-07-23 2014-01-30 Xenon Pharmaceuticals Inc. Cyp8b1 et ses utilisations dans des méthodes thérapeutiques et diagnostiques
DK2895607T3 (da) 2012-09-12 2021-05-25 Quark Pharmaceuticals Inc Dobbeltstrengede oligonukleotidmolekyler til ddit4 og fremgangsmåder til anvendelse deraf
AU2013315524B2 (en) 2012-09-12 2019-01-31 Quark Pharmaceuticals, Inc. Double-stranded oligonucleotide molecules to p53 and methods of use thereof
KR20150060724A (ko) 2012-09-28 2015-06-03 머크 샤프 앤드 돔 코포레이션 Erk 억제제인 신규 화합물
DK2925888T3 (en) 2012-11-28 2017-12-18 Merck Sharp & Dohme COMPOSITIONS AND METHODS OF CANCER TREATMENT
KR102096014B1 (ko) 2012-12-05 2020-04-03 알닐람 파마슈티칼스 인코포레이티드 PCSK9 iRNA 조성물 및 그 사용 방법
JP2016502858A (ja) 2012-12-20 2016-02-01 サレプタ セラピューティクス， インコーポレイテッド 筋ジストロフィを処置するための改善されたエキソンスキッピング組成物
EP2935263B1 (fr) 2012-12-20 2018-12-05 Merck Sharp & Dohme Corp. Imidazopyridines substituées en tant qu'inhibiteurs d'hdm2
EP3263559A1 (fr) 2013-01-22 2018-01-03 Technische Universität Graz Inhibiteur de la lipase
US9540377B2 (en) 2013-01-30 2017-01-10 Merck Sharp & Dohme Corp. 2,6,7,8 substituted purines as HDM2 inhibitors
US20150366890A1 (en) 2013-02-25 2015-12-24 Trustees Of Boston University Compositions and methods for treating fungal infections
JP2016518812A (ja) 2013-03-14 2016-06-30 ガラパゴス・ナムローゼ・フェンノートシャップＧａｌａｐａｇｏｓ Ｎ．Ｖ． 線維症治療において有用な分子標的及び前記標的のインヒビター
US20160054304A1 (en) 2013-03-14 2016-02-25 Galapagos Nv Molecular targets and compounds, and methods to identify the same, useful in the treatment of fibrotic diseases
EA202090946A3 (ru) 2013-03-14 2021-07-30 Сарепта Терапьютикс, Инк. Композиции, обеспечивающие пропускание экзонов, для лечения мышечной дистрофии
HRP20190382T1 (hr) 2013-03-14 2019-04-19 Sarepta Therapeutics, Inc. Pripravci koji preskaču ekson za liječenje mišićne distrofije
ME03043B (fr) 2013-03-14 2018-10-20 Alnylam Pharmaceuticals Inc Compositions d'arni du constituant c5 du complément et leurs procédés d'utilisation
US20160003808A1 (en) 2013-03-14 2016-01-07 Galapagos Nv Molecular targets and compounds, and methods to identify the same, useful in the treatment of diseases associated with epithelial mesenchymal transition
NZ631289A (en) 2013-03-15 2017-08-25 Sarepta Therapeutics Inc Improved compositions for treating muscular dystrophy
US9937231B2 (en) 2013-03-27 2018-04-10 The General Hospital Corporation Methods and agents for treating Alzheimer's disease
TW201515650A (zh) 2013-05-06 2015-05-01 艾爾妮蘭製藥公司 遞送經脂質調配之核酸分子之劑量及方法
US20160186263A1 (en) 2013-05-09 2016-06-30 Trustees Of Boston University Using plexin-a4 as a biomarker and therapeutic target for alzheimer's disease
KR102463973B1 (ko) 2013-05-22 2022-11-07 알닐람 파마슈티칼스 인코포레이티드 ＳＥＲＰＩＮＡ1 ｉＲＮＡ 조성물 및 이의 사용 방법
EA201592225A1 (ru) 2013-05-22 2016-04-29 Элнилэм Фармасьютикалз, Инк. КОМПОЗИЦИИ НА ОСНОВЕ iRNA TMPRSS6 И СПОСОБЫ ИХ ПРИМЕНЕНИЯ
US20160145628A1 (en) 2013-06-18 2016-05-26 Ramot At Tel-Aviv University Ltd. Nanocarrier system for micrornas and uses thereof
EP4039278A1 (fr) 2013-07-11 2022-08-10 Alnylam Pharmaceuticals, Inc. Conjugués d'oligonucléotides et de ligands, et procédé pour leur préparation
WO2015020960A1 (fr) 2013-08-09 2015-02-12 Novartis Ag Nouveaux polynucléotides longs arn non codants (arn lnc)
WO2015034925A1 (fr) 2013-09-03 2015-03-12 Moderna Therapeutics, Inc. Polynucléotides circulaires
EP2853595A1 (fr) 2013-09-30 2015-04-01 Soluventis GmbH Molécules d'ARNsi spécifiques de NOTCH 1
CA2925107A1 (fr) 2013-10-02 2015-04-09 Alnylam Pharmaceuticals, Inc. Compositions et methodes d'inhibition de l'expression du gene lect2
SG11201602631XA (en) 2013-10-04 2016-05-30 Alnylam Pharmaceuticals Inc Compositions and methods for inhibiting expression of the alas1 gene
EP3054959B1 (fr) 2013-10-08 2020-01-08 Promedior Inc. Méthodes de traitement de cancers fibreux
CA2931090A1 (fr) 2013-12-12 2015-06-18 Alnylam Pharmaceuticals, Inc. Composition d'arni d'element de complement et procedes pour les utiliser
SG11201606239UA (en) 2014-01-28 2016-08-30 Buck Inst For Res On Aging Methods and compositions for killing senescent cells and for treating senescence-associated diseases and disorders
AU2015217301A1 (en) 2014-02-11 2016-08-25 Alnylam Pharmaceuticals, Inc. Ketohexokinase (KHK) iRNA compositions and methods of use thereof
WO2015123496A1 (fr) 2014-02-14 2015-08-20 Immune Design Corp. Immunothérapie du cancer par combinaison de stimulation immunitaire locale et systémique
WO2015175510A1 (fr) 2014-05-12 2015-11-19 Alnylam Pharmaceuticals, Inc. Méthodes et compositions pour le traitement d'un trouble associé à serpinc1
BR122020023687B1 (pt) 2014-05-22 2023-03-07 Alnylam Pharmaceuticals, Inc Agente de ácido ribonucleico (rnai) de fita dupla, seus usos e composição farmacêutica
KR20170032373A (ko) 2014-07-15 2017-03-22 이뮨 디자인 코포레이션 Tlr4 작용제 보조제 및 렌티바이러스 벡터를 이용한 프라임-부스트 요법
SG10201806064XA (en) 2014-08-27 2018-08-30 Peptimed Inc Anti-tumor compositions and methods
EP3191591A1 (fr) 2014-09-12 2017-07-19 Alnylam Pharmaceuticals, Inc. Agents polynucléotidiques ciblant le composant du complément c5 et leurs méthodes d'utilisation
KR102473092B1 (ko) 2014-09-15 2022-12-01 칠드런'즈 메디컬 센터 코포레이션 히스톤 h3-리신 트리메틸화를 제거함으로써 체세포 핵 이식(scnt) 효율을 증가시키는 방법 및 조성물
TWI864340B (zh) 2014-10-10 2024-12-01 美商艾爾妮蘭製藥公司 用於抑制hao1(羥酸氧化酶1(乙醇酸鹽氧化酶))基因表現的組合物及方法
EP3207138B1 (fr) 2014-10-17 2020-07-15 Alnylam Pharmaceuticals, Inc. Agents polynucléotidiques de ciblage d'acide aminolévulinique synthase-1 (alas1) et utilisations de ceux-ci
EP3904519A1 (fr) 2014-10-30 2021-11-03 Genzyme Corporation Agents polynucléotidiques ciblant serpinc 1 (at3) et leurs méthodes d'utilisation
SG11201703638XA (en) 2014-11-10 2017-06-29 Alnylam Pharmaceuticals Inc Hepatitis b virus (hbv) irna compositions and methods of use thereof
CA2968114A1 (fr) 2014-11-17 2016-05-26 Alnylam Pharmaceuticals, Inc. Compositions d'arni d'apolipoproteine c3 (apoc3) et procedes d'utilisation de ces compositions
EP3224362B1 (fr) 2014-11-26 2024-09-25 The Regents of The University of California Compositions thérapeutiques comprenant des facteurs de transcription et leurs procédés de préparation et d'utilisation
WO2016083624A1 (fr) 2014-11-28 2016-06-02 Silence Therapeutics Gmbh Moyens d'inhibition de l'expression d'edn1
WO2016130806A2 (fr) 2015-02-13 2016-08-18 Alnylam Pharmaceuticals, Inc. Compositions d'arni du gène codant pour la protéine 3 contenant un domaine phospholipase de type patatine (pnpla3) et leurs procédés d'utilisation
WO2016142948A1 (fr) 2015-03-11 2016-09-15 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Oligonucléotides leurres pour le traitement de maladies
MX2017012610A (es) 2015-04-08 2018-03-16 Alnylam Pharmaceuticals Inc Composiciones y metodos para inhibir la expresion del gen lect2.
WO2016201301A1 (fr) 2015-06-12 2016-12-15 Alnylam Pharmaceuticals, Inc. Compositions d'arni de composant du complément c5 et leurs procédés d'utilisation
EP3310918B1 (fr) 2015-06-18 2020-08-05 Alnylam Pharmaceuticals, Inc. Agents polynucléotidiques ciblant l'hydroxyacide oxydase (glycolate oxydase, hao1) et procédés d'utilisation de ceux-ci
WO2016209862A1 (fr) 2015-06-23 2016-12-29 Alnylam Pharmaceuticals, Inc. Compositions d'arndb contre un gène de glucokinase (gck) et leurs procédés d'utilisation
US10494632B2 (en) 2015-07-10 2019-12-03 Alnylam Pharmaceuticals, Inc. Insulin-like growth factor binding protein, acid labile subunit (IGFALS) compositions and methods of use thereof
DK3324980T3 (da) 2015-07-17 2022-02-14 Alnylam Pharmaceuticals Inc Multimålrettede enkeltenhedskonjugater
CA2994719A1 (fr) 2015-08-21 2017-03-02 The Children's Hospital Of Philadelphia Procedes de traitement de conditions auto-immunes chez des patients comprenant des variations genetiques dans dcr3 ou dans un gene de reseau de dcr3
CN108368507B (zh) 2015-09-02 2022-03-22 阿尔尼拉姆医药品有限公司 程序性细胞死亡1配体1(PD-L1)的iRNA组合物及其使用方法
US20190054113A1 (en) 2015-09-30 2019-02-21 Sarepta Therapeutics, Inc. Methods for treating muscular dystrophy
US12241053B2 (en) 2015-10-09 2025-03-04 The Brigham And Women's Hospital, Inc. Modulation of novel immune checkpoint targets
EP3368689B1 (fr) 2015-10-28 2020-06-17 The Broad Institute, Inc. Compositions d'évaluation et de modulation des réponses immunitaires à l'aide de signatures génétiques de cellules immunitaires
WO2017075465A1 (fr) 2015-10-28 2017-05-04 The Broad Institute Inc. Compositions et procédés d'évaluation et de modulation des réponses immunitaires par détection et ciblage de gata3
WO2017075451A1 (fr) 2015-10-28 2017-05-04 The Broad Institute Inc. Compositions et procédés d'évaluation et de modulation des réponses immunitaires par détection et ciblage de pou2af1
SG11201804729RA (en) 2015-12-07 2018-07-30 Genzyme Corp Methods and compositions for treating a serpinc1-associated disorder
WO2017184586A1 (fr) 2016-04-18 2017-10-26 The Trustees Of Columbia University In The City Of New York Cibles thérapeutiques impliquées dans la progression de la stéatose hépatique non alcoolique (nash)
WO2017184689A1 (fr) 2016-04-19 2017-10-26 Alnylam Pharmaceuticals, Inc. Composition d'arni de protéine de liaison de lipoprotéines haute densité (hdlbp/vigiline) et procédés pour les utiliser
WO2017190041A1 (fr) 2016-04-29 2017-11-02 Sarepta Therapeutics, Inc. Analogues d'oligonucléotide ciblant lmna humain
WO2017214518A1 (fr) 2016-06-10 2017-12-14 Alnylam Pharmaceuticals, Inc. Compositions d'arni du composant c5 du complément et leurs méthodes d'utilisation pour traiter l'hémoglobinurie paroxystique nocturne (hpn)
AU2017290231A1 (en) 2016-06-30 2019-02-07 Sarepta Therapeutics, Inc. Exon skipping oligomers for muscular dystrophy
EP3269734A1 (fr) 2016-07-15 2018-01-17 Fundació Privada Institut d'Investigació Oncològica de Vall-Hebron Procédés et compositions pour le traitement du cancer
US20190262399A1 (en) 2016-09-07 2019-08-29 The Broad Institute, Inc. Compositions and methods for evaluating and modulating immune responses
WO2018067991A1 (fr) 2016-10-07 2018-04-12 The Brigham And Women's Hospital, Inc. Modulation de nouvelles cibles de points de contrôle immunitaires
WO2018071283A1 (fr) 2016-10-12 2018-04-19 Merck Sharp & Dohme Corp. Inhibiteurs de kdm5
TW202313978A (zh) 2016-11-23 2023-04-01 美商阿尼拉製藥公司 絲胺酸蛋白酶抑制因子Ａ１ ｉＲＮＡ組成物及其使用方法
EP3330276A1 (fr) 2016-11-30 2018-06-06 Universität Bern Nouveaux nucléosides bicycliques et oligomères préparés à partir de ces derniers
AU2017376950B2 (en) 2016-12-16 2024-02-22 Alnylam Pharmaceuticals, Inc. Methods for treating or preventing TTR-associated diseases using transthyretin (TTR) iRNA compositions
EP3554552B1 (fr) 2016-12-19 2022-08-17 Sarepta Therapeutics, Inc. Conjugués oligomères de sauts d'exons pour la dystrophie musculaire
LT3554553T (lt) 2016-12-19 2022-08-25 Sarepta Therapeutics, Inc. Egzoną šalinantys oligomero konjugatai nuo raumenų distrofojos
CN117298290A (zh) 2016-12-19 2023-12-29 萨勒普塔医疗公司 用于肌肉萎缩症的外显子跳跃寡聚体缀合物
SG11201909572QA (en) 2017-04-18 2019-11-28 Alnylam Pharmaceuticals Inc Methods for the treatment of subjects having a hepatitis b virus (hbv) infection
JP2020530442A (ja) 2017-07-10 2020-10-22 ジェンザイム・コーポレーション 血友病を有する対象の出血事象を処置するための方法および組成物
CA3069868A1 (fr) 2017-07-13 2019-01-17 Alnylam Pharmaceuticals Inc. Compositions d'arni de lactate deshydrogenase a (ldha) et leurs procedes d'utilisation
EA201991450A1 (ru) 2017-09-22 2019-12-30 Сарепта Терапьютикс, Инк. Конъюгаты олигомеров для пропуска экзона при мышечной дистрофии
JP2020536057A (ja) 2017-09-28 2020-12-10 サレプタ セラピューティクス， インコーポレイテッド 筋ジストロフィーを処置するための併用療法
JP2020536060A (ja) 2017-09-28 2020-12-10 サレプタ セラピューティクス， インコーポレイテッド 筋ジストロフィーを処置するための併用療法
EP3687577A1 (fr) 2017-09-28 2020-08-05 Sarepta Therapeutics, Inc. Polythérapies pour traiter une dystrophie musculaire
US11866701B2 (en) 2017-11-01 2024-01-09 Alnylam Pharmaceuticals, Inc. Complement component C3 iRNA compositions and methods of use thereof
WO2019094311A1 (fr) 2017-11-08 2019-05-16 Merck Sharp & Dohme Corp. Inhibiteurs de prmt5
US11098059B2 (en) 2017-11-08 2021-08-24 Merck Sharp & Dohme Corp. PRMT5 inhibitors
US20200385719A1 (en) 2017-11-16 2020-12-10 Alnylam Pharmaceuticals, Inc. Kisspeptin 1 (kiss1) irna compositions and methods of use thereof
EP3714054A1 (fr) 2017-11-20 2020-09-30 Alnylam Pharmaceuticals, Inc. Compositions d'arni de composant p amyloïde serique et leurs procédés d'utilisation
WO2019126097A1 (fr) 2017-12-18 2019-06-27 Alnylam Pharmaceuticals, Inc. Compositions d'arni à boîte 1 du groupe de haute mobilité (hmgb1) et leurs procédés d'utilisation
WO2019170731A1 (fr) 2018-03-07 2019-09-12 Sanofi Précurseurs nucléotidiques, analogues nucléotidiques et composés oligomères les contenant
TWI851574B (zh) 2018-05-14 2024-08-11 美商阿尼拉製藥公司 血管收縮素原（ＡＧＴ）ｉＲＮＡ組成物及其使用方法
US10758629B2 (en) 2018-05-29 2020-09-01 Sarepta Therapeutics, Inc. Exon skipping oligomer conjugates for muscular dystrophy
EP3806868A4 (fr) 2018-06-13 2022-06-22 Sarepta Therapeutics, Inc. Oligomères induisant un saut d'exon pour la dystrophie musculaire
TW202020153A (zh) 2018-07-27 2020-06-01 美商薩羅塔治療公司 用於肌肉萎縮症之外顯子跳躍寡聚物
EP3833667B1 (fr) 2018-08-07 2024-03-13 Merck Sharp & Dohme LLC Inhibiteurs de prmt5
BR112021002267A8 (pt) 2018-08-07 2023-02-07 Merck Sharp & Dohme Inibidores de prmt5
EP3833668B1 (fr) 2018-08-07 2025-03-19 Merck Sharp & Dohme LLC Inhibiteurs de prmt5
MA70658B1 (fr) 2018-08-13 2026-04-30 Alnylam Pharmaceuticals, Inc. Compositions d'agent d'arndb du virus de l'hépatite b (vhb) et leurs méthodes d'utilisation
US11987792B2 (en) 2018-08-16 2024-05-21 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of the LECT2 gene
WO2020060986A1 (fr) 2018-09-18 2020-03-26 Alnylam Pharmaceuticals, Inc. Compositions d'arni de cétohexokinase (khk) et leurs procédés d'utilisation
US10913951B2 (en) 2018-10-31 2021-02-09 University of Pittsburgh—of the Commonwealth System of Higher Education Silencing of HNF4A-P2 isoforms with siRNA to improve hepatocyte function in liver failure
JP7707065B2 (ja) 2018-11-23 2025-07-14 サノフイ Ａｎｇｐｔｌ８を阻害するための新規のｒｎａ組成物および方法
WO2020117706A1 (fr) 2018-12-03 2020-06-11 Triplet Therapeutics, Inc. Méthodes thérapeutiques pour les maladies par expansion de répétitions trinucléotidiques associés à une activité mlh3
IL321667A (en) 2018-12-13 2025-08-01 Sarepta Therapeutics Inc Exon skipping oligomer conjugates for muscular dystrophy
CN113631709A (zh) 2018-12-20 2021-11-09 普拉克西斯精密药物股份有限公司 用于治疗kcnt1相关病症的组合物和方法
LT4285929T (lt) 2018-12-20 2026-01-12 Humabs Biomed Sa Kompleksinė hbv terapija
WO2020187998A1 (fr) 2019-03-19 2020-09-24 Fundació Privada Institut D'investigació Oncològica De Vall Hebron Polythérapie à l'aide d'omomyc et d'un anticorps se liant à pd-1 ou ctla-4 pour le traitement du cancer
SG11202108757XA (en) 2019-03-28 2021-10-28 Sarepta Therapeutics Inc Methods for treating muscular dystrophy with casimersen
US20220193246A1 (en) 2019-04-18 2022-06-23 Sarepta Therapeutics, Inc. Compositions for treating muscular dystrophy
JP2022536495A (ja) 2019-06-19 2022-08-17 サレプタ セラピューティクス， インコーポレイテッド 筋ジストロフィーを治療する方法
JP2022541197A (ja) 2019-07-16 2022-09-22 ユニヴェルシテ・コート・ダジュール 腫瘍溶解性ウイルスに対する対象の感受性又は抵抗性を評価する方法、組換えウイルス、その調製、及び使用
EP4007812A1 (fr) 2019-08-01 2022-06-08 Alnylam Pharmaceuticals, Inc. Compositions d'arni de la famille des serpines 2 (serpinf2) et leurs procedes d'utilisation
EP4007811A2 (fr) 2019-08-01 2022-06-08 Alnylam Pharmaceuticals, Inc. Compositions d'arni de carboxypeptidase b2 (cpb2) et leurs procédés d'utilisation
WO2021030522A1 (fr) 2019-08-13 2021-02-18 Alnylam Pharmaceuticals, Inc. Compositions d'agent d'arni à sous-unité de protéine ribosomale 25 (rps25) et leurs procédés d'utilisation
ES2918001B2 (es) 2019-08-14 2026-05-13 Acuitas Therapeutics Inc Nanoparticulas lipidicas mejoradas para el suministro de acidos nucleicos
WO2021037972A1 (fr) 2019-08-27 2021-03-04 Sanofi Compositions et procédés d'inhibition de pcsk9
US20220290152A1 (en) 2019-09-03 2022-09-15 Alnylam Pharmaceuticals, Inc. Compositions and methods for inhibiting expression of the lect2 gene
JP7667775B2 (ja) 2019-09-05 2025-04-23 サノフイ ヌクレオチド類似体を含有するオリゴヌクレオチド
CN114391040A (zh) 2019-09-23 2022-04-22 欧米茄治疗公司 用于调节载脂蛋白b(apob)基因表达的组合物和方法
AU2020352552A1 (en) 2019-09-23 2022-03-17 Omega Therapeutics, Inc. Compositions and methods for modulating hepatocyte nuclear factor 4-alpha (HNF4α) gene expression
WO2021067747A1 (fr) 2019-10-04 2021-04-08 Alnylam Pharmaceuticals, Inc. Compositions et méthodes pour le silençage de l'expression génique de l'ugt1a1
US20240050529A1 (en) 2019-10-07 2024-02-15 University Of Virginia Patent Foundation Modulating lymphatic vessels in neurological disease
EP3808763A1 (fr) 2019-10-17 2021-04-21 Fundació Institut d'Investigació Biomèdica de Bellvitge (IDIBELL) Composés d'immunomodulation
WO2021076828A1 (fr) 2019-10-18 2021-04-22 Alnylam Pharmaceuticals, Inc. Compositions d'arni de membre de la famille des transporteurs de solutés et leurs procédés d'utilisation
BR112022007540A2 (pt) 2019-10-22 2022-07-12 Alnylam Pharmaceuticals Inc Composições de irna de componente complementar c3 e métodos de uso das mesmas
WO2021087325A1 (fr) 2019-11-01 2021-05-06 Alnylam Pharmaceuticals, Inc. Compositions et méthodes pour rendre silencieuse l'expression du gène de fusion dnajb1-prkaca
CN114728018B (zh) 2019-11-01 2024-07-19 阿尔尼拉姆医药品有限公司 亨廷顿(HTT)iRNA药剂组合物及其使用方法
TW202526019A (zh) 2019-11-13 2025-07-01 美商阿尼拉製藥公司 用於治療血管收縮素原相關病症之方法及組成物
EP4061945A1 (fr) 2019-11-22 2022-09-28 Alnylam Pharmaceuticals, Inc. Compositions d'agent arni d'ataxine 3 (atxn3) et leurs procédés d'utilisation
TWI897902B (zh) 2019-12-13 2025-09-21 美商阿尼拉製藥公司 人類染色體9開讀框72 (C9ORF72) iRNA劑組成物及其使用方法
WO2021126734A1 (fr) 2019-12-16 2021-06-24 Alnylam Pharmaceuticals, Inc. Compositions d'arni du gène codant pour la protéine 3 contenant un domaine phospholipase de type patatine (pnpla3) et leurs méthodes d'utilisation
EP4706689A2 (fr) 2019-12-19 2026-03-11 Entrada Therapeutics, Inc. Compositions pour administrer des composés antisens
EP4093426A1 (fr) 2020-01-24 2022-11-30 University of Virginia Patent Foundation Modulation de vaisseaux lymphatiques dans une maladie neurologique
WO2021154941A1 (fr) 2020-01-31 2021-08-05 Alnylam Pharmaceuticals, Inc. Compositions d'arni du composant c5 du complément destinées à être utilisées dans le traitement de la sclérose latérale amyotrophique (sla)
MX2022009763A (es) 2020-02-10 2022-09-09 Alnylam Pharmaceuticals Inc Composiciones y metodos para silenciar la expresion del factor de crecimiento endotelial vascular a (vegf-a).
WO2021167841A1 (fr) 2020-02-18 2021-08-26 Alnylam Pharmaceuticals, Inc. Compositions d'arni d'apolipoprotéine c3 (apoc3) et leurs méthodes d'utilisation
WO2021178736A1 (fr) 2020-03-06 2021-09-10 Alnylam Pharmaceuticals, Inc. Compositions d'arni de cétohexokinase (khk) et leurs procédés d'utilisation
JP7802676B2 (ja) 2020-03-06 2026-01-20 アルナイラム ファーマシューティカルズ， インコーポレイテッド トランスサイレチン（ｔｔｒ）の発現を阻害するための組成物および方法
WO2021183720A1 (fr) 2020-03-11 2021-09-16 Omega Therapeutics, Inc. Compositions et procédés de modulation de l'expression génique de forkhead box p3 (foxp3)
EP4121018A4 (fr) 2020-03-17 2024-07-03 Genevant Sciences Gmbh Lipides cationiques pour l'administration de nanoparticules lipidiques d'agents thérapeutiques à des cellules stellaires hépatiques
EP4121534A1 (fr) 2020-03-18 2023-01-25 Alnylam Pharmaceuticals, Inc. Compositions et méthodes pour traiter des sujets ayant un variant de gène d'alanine-glyoxylate aminotransférase hétérozygote (agxt)
WO2021195307A1 (fr) 2020-03-26 2021-09-30 Alnylam Pharmaceuticals, Inc. Compositions d'arni de coronavirus et leurs procédés d'utilisation
EP4127171A2 (fr) 2020-03-30 2023-02-08 Alnylam Pharmaceuticals, Inc. Compositions et procédés de silençage de l'expression du gène dnajc15
US12534731B2 (en) 2020-04-01 2026-01-27 Alnylam Pharmaceuticals, Inc. Alpha-2A adrenergic receptor (ADRA2A) iRNA agent compositions and methods of use thereof
CA3179411A1 (fr) 2020-04-06 2021-10-14 Alnylam Pharmaceuticals, Inc. Compositions et methodes pour le silencage de l'expression de myoc
WO2021206922A1 (fr) 2020-04-07 2021-10-14 Alnylam Pharmaceuticals, Inc. Compositions d'arni de la serine protease 2 transmembranaire (tmprss2) et leurs procédés d'utilisation
IL297130A (en) 2020-04-07 2022-12-01 Alnylam Pharmaceuticals Inc Compositions and methods for silencing scn9a expression
WO2021206917A1 (fr) 2020-04-07 2021-10-14 Alnylam Pharmaceuticals, Inc. Compositions arni d'enzyme de conversion de l'angiotensine 2 (eca2) et procédés d'utilisation associés
WO2021222065A1 (fr) 2020-04-27 2021-11-04 Alnylam Pharmaceuticals, Inc. Compositions d'agent d'arni de l'apolipoprotéine e (apoe) et leurs procédés d'utilisation
EP4142738A1 (fr) 2020-04-30 2023-03-08 Alnylam Pharmaceuticals, Inc. Compositions d'arni du facteur b du complément (cfb) et leurs procédés d'utilisation
WO2021231692A1 (fr) 2020-05-15 2021-11-18 Korro Bio, Inc. Procédés et compositions pour l'édition médiée par adar d'otoferline (otof)
WO2021231679A1 (fr) 2020-05-15 2021-11-18 Korro Bio, Inc. Procédés et compositions pour l'édition médiée par adar de la protéine bêta 2 de jonction lacunaire (gjb2)
WO2021231680A1 (fr) 2020-05-15 2021-11-18 Korro Bio, Inc. Procédés et compositions pour l'édition médiée par adar de la protéine 2 de liaison méthyl-cpg (mecp2)
EP4150088A1 (fr) 2020-05-15 2023-03-22 Korro Bio, Inc. Procédés et compositions pour l'édition médiée par adar d'argininosuccinate synthétase (ass1)
WO2021231691A1 (fr) 2020-05-15 2021-11-18 Korro Bio, Inc. Procédés et compositions pour l'édition médiée par adar de rétinoschisine 1 (rs1)
EP4150077A1 (fr) 2020-05-15 2023-03-22 Korro Bio, Inc. Procédés et compositions pour l'édition médiée par adar de la protéine 1 de type canal transmembranaire (tmc1)
WO2021231673A1 (fr) 2020-05-15 2021-11-18 Korro Bio, Inc. Procédés et compositions pour l'édition médiée par adar de la kinase 2 à répétition riche en leucine (lrrk2)
WO2021231698A1 (fr) 2020-05-15 2021-11-18 Korro Bio, Inc. Procédés et compositions pour l'édition médiée par adar d'argininosuccinate lyase (asl)
EP4153746A1 (fr) 2020-05-21 2023-03-29 Alnylam Pharmaceuticals, Inc. Compositions et procédés d'inhibition de l'expression du gène marc1
US11408000B2 (en) 2020-06-03 2022-08-09 Triplet Therapeutics, Inc. Oligonucleotides for the treatment of nucleotide repeat expansion disorders associated with MSH3 activity
US20230203496A1 (en) 2020-06-09 2023-06-29 Alnylam Pharmaceuticals, Inc. Sirna compositions and methods for silencing gpam (glycerol-3-phosphate acyltransferase 1, mitochondrial) expression
AU2021292296A1 (en) 2020-06-18 2023-01-19 Alnylam Pharmaceuticals, Inc. Xanthine dehydrogenase (XDH) iRNA compositions and methods of use thereof
AR122722A1 (es) 2020-06-24 2022-09-28 Vir Biotechnology Inc Anticuerpos que neutralizan el virus de la hepatitis b y sus usos
EP4217489A1 (fr) 2020-09-24 2023-08-02 Alnylam Pharmaceuticals, Inc. Compositions d'arni de dipeptidyle peptidase 4 (dpp4) et leurs procédés d'utilisation
TW202229552A (zh) 2020-10-05 2022-08-01 美商艾拉倫製藥股份有限公司 Ｇ蛋白－偶合受體７５（ＧＰＲ７５）ｉＲＮＡ組成物及其使用方法
EP4229200A1 (fr) 2020-10-16 2023-08-23 Sanofi Nouvelles compositions d'arn et méthodes d'inhibition d'angptl3
JP2023545502A (ja) 2020-10-16 2023-10-30 サノフイ リポタンパク質（ａ）を阻害するためのｒｎａ組成物および方法
CN116368146A (zh) 2020-10-20 2023-06-30 赛诺菲 脱唾液酸糖蛋白受体的新型配体
AU2021365822A1 (en) 2020-10-21 2023-06-08 Alnylam Pharmaceuticals, Inc. Methods and compositions for treating primary hyperoxaluria
WO2022087329A1 (fr) 2020-10-23 2022-04-28 Alnylam Pharmaceuticals, Inc. Compositions d'arni de la mucine 5b (muc5b) et leurs méthodes d'utilisation
WO2022103999A1 (fr) 2020-11-13 2022-05-19 Alnylam Pharmaceuticals, Inc. Compositions d'arni du facteur de coagulation v (f5) et leurs procédés d'utilisation
EP4256053A1 (fr) 2020-12-01 2023-10-11 Alnylam Pharmaceuticals, Inc. Méthodes et compositions pour l'inhibition de l'expression du gène de l'hao1 (hydroxyacide oxydase 1 (glycolate oxydase))
WO2022125490A1 (fr) 2020-12-08 2022-06-16 Alnylam Pharmaceuticals, Inc. Compositions d'arni du facteur de coagulation x (f10) et leurs méthodes d'utilisation
CN117479963A (zh) 2020-12-18 2024-01-30 盖纳万科学有限公司 Peg脂质和脂质纳米粒子
EP4274896A1 (fr) 2021-01-05 2023-11-15 Alnylam Pharmaceuticals, Inc. Compositions d'arni du composant du complément 9 (c9) et leurs méthodes d'utilisation
EP4291654A2 (fr) 2021-02-12 2023-12-20 Alnylam Pharmaceuticals, Inc. Compositions d'arni de superoxyde dismutase 1 (sod1) et procédés d'utilisation correspondants pour traiter ou prévenir des maladies neurodégénératives associées à la superoxyde dismutase 1 (sod1)
WO2022182864A1 (fr) 2021-02-25 2022-09-01 Alnylam Pharmaceuticals, Inc. Compositions à base d'arni de protéine prion (prnp) et procédés et procédés d'utilisation de celles-ci
JP2024515423A (ja) 2021-02-26 2024-04-10 アルナイラム ファーマシューティカルズ， インコーポレイテッド ケトヘキソキナーゼ（ＫＨＫ）ｉＲＮＡ組成物およびその使用方法
AU2022231003A1 (en) 2021-03-04 2023-09-14 Alnylam Pharmaceuticals, Inc. Angiopoietin-like 3 (angptl3) irna compositions and methods of use thereof
EP4305169A1 (fr) 2021-03-12 2024-01-17 Alnylam Pharmaceuticals, Inc. Compositions d'arni de la glycogène synthase kinase 3 alpha (gsk3a) et leurs procédés d'utilisation
IL307239A (en) 2021-03-29 2023-11-01 Alnylam Pharmaceuticals Inc Preparations containing Huntingtin IRNA factor (HTT) and methods of using them
US20250243244A1 (en) 2021-03-31 2025-07-31 Entrada Therapeutics, Inc. Cyclic cell penetrating peptides
WO2022212153A1 (fr) 2021-04-01 2022-10-06 Alnylam Pharmaceuticals, Inc. Compositions d'arni de proline déshydrogénase 2 (prodh2) et procédés d'utilisation associés
BR112023022284A2 (pt) 2021-04-26 2023-12-26 Alnylam Pharmaceuticals Inc Composições de irna de protease transmembrana, serina 6 (tmprss6) e métodos de uso da mesma
EP4330396A1 (fr) 2021-04-29 2024-03-06 Alnylam Pharmaceuticals, Inc. Transducteur de signal et activateur de compositions d'arni du facteur de transcription 6 (stat6) et procédés d'utilisation correspondants
CA3216839A1 (fr) 2021-04-30 2022-11-03 Sarepta Therapeutics, Inc. Procedes de traitement pour dystrophie musculaire
EP4337261A2 (fr) 2021-05-10 2024-03-20 Entrada Therapeutics, Inc. Compositions et procédés de modulation de l'épissage d'arnm
WO2022240721A1 (fr) 2021-05-10 2022-11-17 Entrada Therapeutics, Inc. Compositions et méthodes de modulation de l'activité du facteur 5 de régulation de l'interféron (irf-5)
AU2022271873A1 (en) 2021-05-10 2024-01-04 Entrada Therapeutics, Inc. Compositions and methods for intracellular therapeutics
EP4341401A1 (fr) 2021-05-18 2024-03-27 Alnylam Pharmaceuticals, Inc. Compositions d'arni de cotransporteur-2 de sodium-glucose (sglt2) et leurs procédés d'utilisation
EP4341405A1 (fr) 2021-05-20 2024-03-27 Korro Bio, Inc. Procédés et compositions pour l'édition médiée par adar
WO2022256283A2 (fr) 2021-06-01 2022-12-08 Korro Bio, Inc. Méthodes de restauration de la fonction protéique par adar
JP2024522996A (ja) 2021-06-02 2024-06-25 アルナイラム ファーマシューティカルズ， インコーポレイテッド パタチン様ホスホリパーゼドメイン含有３（ＰＮＰＬＡ３）ｉＲＮＡ組成物およびその使用方法
BR112023025224A2 (pt) 2021-06-04 2024-02-27 Alnylam Pharmaceuticals Inc Quadro de leitura aberto 72 do cromossomo humano 9 (c9orf72) composições de agente de irna e métodos de uso dos mesmos
EP4351541A2 (fr) 2021-06-08 2024-04-17 Alnylam Pharmaceuticals, Inc. Compositions et méthodes de traitement ou de prévention de la maladie de stargardt et/ou de troubles associés à la protéine 4 de liaison du rétinol (rbp4)
GEAP202416440A (en) 2021-06-23 2024-04-25 Entrada Therapeutics Inc Antisense compounds and methods for targeting cug repeats
US20230194709A9 (en) 2021-06-29 2023-06-22 Seagate Technology Llc Range information detection using coherent pulse sets with selected waveform characteristics
US20250034564A1 (en) 2021-06-29 2025-01-30 Korro Bio, Inc. Methods and Compositions for ADAR-Mediated Editing
MX2023015489A (es) 2021-06-30 2024-01-19 Alnylam Pharmaceuticals Inc Metodos y composiciones para tratar un trastorno asociado al angiotensinogeno (agt).
WO2023283359A2 (fr) 2021-07-07 2023-01-12 Omega Therapeutics, Inc. Compositions et procédés de modulation de l'expression génique de la protéine 1 du récepteur frizzled secrété (sfrp1)
WO2023283403A2 (fr) 2021-07-09 2023-01-12 Alnylam Pharmaceuticals, Inc. Composés bis-arni pour administration au snc
WO2023003805A1 (fr) 2021-07-19 2023-01-26 Alnylam Pharmaceuticals, Inc. Méthodes et compositions pour traiter des sujets ayant ou ayant un risque de développer une maladie ou un trouble d'hyperoxalurie non primaire
KR20240036041A (ko) 2021-07-21 2024-03-19 알닐람 파마슈티칼스 인코포레이티드 대사 장애-연관 표적 유전자 iRNA 조성물 및 이의 사용 방법
JP2024528701A (ja) 2021-07-23 2024-07-30 アルナイラム ファーマシューティカルズ， インコーポレイテッド ベータカテニン（ＣＴＮＮＢ１）ｉＲＮＡ組成物およびその使用方法
JP2024529437A (ja) 2021-07-29 2024-08-06 アルナイラム ファーマシューティカルズ， インコーポレイテッド ３－ヒドロキシ－３－メチルグリタル－ＣＯＡレダクターゼ（ＨＭＧＣＲ）ｉＲＮＡ組成物およびその使用方法
TW202328445A (zh) 2021-08-03 2023-07-16 美商艾拉倫製藥股份有限公司 甲狀腺素運載蛋白（ＴＴＲ）ｉＲＮＡ組成物及其使用方法
PE20241132A1 (es) 2021-08-04 2024-05-24 Alnylam Pharmaceuticals Inc Composiciones de arni y metodos para silenciar el angiotensinogeno (agt)
US12465647B2 (en) 2021-08-05 2025-11-11 Sanegene Bio Usa Inc. 1′-alkyl modified ribose derivatives and methods of use
IL310407A (en) 2021-08-13 2024-03-01 Alnylam Pharmaceuticals Inc Factor XII (F12) IRNA compositions and methods of using them
MX2024002440A (es) 2021-08-31 2024-03-08 Alnylam Pharmaceuticals Inc Composiciones de acido ribonucleico de interferencia (arni) del efector b similar al factor de fragmentacion de adn subunidad alfa (dffa) que induce la muerte celular (cideb) y metodos de uso de estas.
JP2024535850A (ja) 2021-09-17 2024-10-02 アルナイラム ファーマシューティカルズ， インコーポレイテッド 補体成分（Ｃ３）をサイレンシングするためのｉＲＮＡ組成物および方法
EP4405478A1 (fr) 2021-09-20 2024-07-31 Alnylam Pharmaceuticals, Inc. Compositions modulatrices de la sous-unité beta e de l'inhibine (inhbe) et leurs procédés d'utilisation
AU2022352676A1 (en) 2021-09-22 2024-03-14 Sanegene Bio Usa Inc. 2'-alkyl or 3'- alkyl modified ribose derivatives for use in the in-vivo delivery of oligonucleotides
MX2024003519A (es) 2021-09-24 2024-04-01 Alnylam Pharmaceuticals Inc Composiciones de agentes de acido ribonucleico de interferencia (arni) de proteina tau asociada a microtubulos (mapt) y sus metodos de uso.
JP2024542929A (ja) 2021-10-05 2024-11-19 セーンジーン バイオ ユーエスエー インコーポレイティド ポリヒドロキシル化されたシクロペンタン誘導体及び使用方法
IL311864A (en) 2021-10-15 2024-06-01 Alnylam Pharmaceuticals Inc IRNA compositions for extrahepatic administration and methods of using them
AU2022368907A1 (en) 2021-10-20 2024-05-02 University Of Copenhagen Rejuvenation treatment of age-related white matter loss
AU2022370009A1 (en) 2021-10-22 2024-05-16 Korro Bio, Inc. Methods and compositions for disrupting nrf2-keap1 protein interaction by adar mediated rna editing
TW202334418A (zh) 2021-10-29 2023-09-01 美商艾拉倫製藥股份有限公司 杭丁頓（ＨＴＴ）ｉＲＮＡ劑組成物及其使用方法
CA3234636A1 (fr) 2021-10-29 2023-05-04 Alnylam Pharmaceuticals, Inc. Compositions d'arni du facteur b du complement (cfb) et leurs procedes d'utilisation
US20250011767A1 (en) 2021-11-18 2025-01-09 Circularis Biotechnologies, Inc. Compositions and methods for production of circular nucleic acid molecules
KR20240126870A (ko) 2021-12-22 2024-08-21 캠프4 테라퓨틱스 코포레이션 조절 rna들을 표적으로 하는 안티센스 올리고뉴클레오티드를 사용한 유전자 전사의 조절
EP4469575A2 (fr) 2022-01-24 2024-12-04 Alnylam Pharmaceuticals, Inc. Compositions d'agent d'arni d'enzyme de voie de biosynthèse de sulfate d'héparine et leurs méthodes d'utilisation
CN118922401A (zh) 2022-01-31 2024-11-08 盖纳万科学有限公司 用于脂质纳米颗粒的可电离阳离子脂质
CN118900701A (zh) 2022-01-31 2024-11-05 盖纳万科学有限公司 聚(烷基噁唑啉)-脂质缀合物及含有聚(烷基噁唑啉)-脂质缀合物的脂质颗粒
JP2025508785A (ja) 2022-02-22 2025-04-10 セーンジーン バイオ ユーエスエー インコーポレイティド ５’修飾炭素環式リボヌクレオチド誘導体及び使用方法
JP2025509438A (ja) 2022-03-17 2025-04-11 サレプタ セラピューティクス， インコーポレイテッド ホスホロジアミデートモルホリノオリゴマーコンジュゲート
AU2023283551A1 (en) 2022-06-10 2024-12-19 Camp4 Therapeutics Corporation Methods of modulating progranulin expression using antisense oligonucleotides targeting regulatory rnas
WO2024015796A1 (fr) 2022-07-11 2024-01-18 Sanegene Bio Usa Inc. Dérivés du ribose modifié en 2' optimisés et procédés d'utilisation
JP2025527531A (ja) 2022-08-18 2025-08-22 アルナイラム ファーマシューティカルズ， インコーポレイテッド ユニバーサル非標的ｓｉｒｎａ組成物およびその使用方法
TW202424193A (zh) 2022-09-15 2024-06-16 美商艾拉倫製藥股份有限公司 第１３型１７β－羥基類固醇去氫酶（ＨＳＤ１７Ｂ１３）ｉＲＮＡ組成物及其使用方法
AU2023370106A1 (en) 2022-10-25 2025-05-08 Fundació Privada Institut D'investigació Oncològica De Vall Hebron Combination therapy for the treatment of cancer
EP4612292A1 (fr) 2022-11-02 2025-09-10 Sarepta Therapeutics, Inc. Formulation d'un conjugué oligomère antisens
JP2025540101A (ja) 2022-12-01 2025-12-11 キャンプ４ セラピューティクス コーポレイション 調節ｒｎａを標的とするアンチセンスオリゴヌクレオチドを用いたｓｙｎｇａｐ１遺伝子転写の調節
EP4658294A2 (fr) 2023-02-02 2025-12-10 University of Rochester Remplacement compétitif de cellules gliales
EP4410825A1 (fr) 2023-02-03 2024-08-07 Servizo Galego de Saude Fragments du domaine n-terminal de gsdmb pour le traitement du cancer
AU2024217843A1 (en) 2023-02-09 2025-07-31 Alnylam Pharmaceuticals, Inc. Reversir molecules and methods of use thereof
KR20260005960A (ko) 2023-05-03 2026-01-12 사네진 바이오 유에스에이 인크. 전신, 중추 신경계, 말초 신경계, 및 안구 전달을 위한 지질 기반 접합체
WO2024243438A2 (fr) 2023-05-23 2024-11-28 Omega Therapeutics, Inc. Compositions et procédés de réduction de l'expression du gène cxcl9, cxcl10 et cxcl11
WO2024251846A1 (fr) 2023-06-07 2024-12-12 Fundació Privada Institut D'investigació Oncològica De Vall Hebron Polythérapie avec des inhibiteurs de mek pour le traitement du cancer
EP4473974A1 (fr) 2023-06-07 2024-12-11 Peptomyc, S.L. Polythérapie avec omomyc et des inhibiteurs de kras pour le traitement du cancer
KR20260020418A (ko) 2023-06-07 2026-02-11 펀다시오 프리바다 인스티튜트 드인베스티가시오 온콜로지카 데 발 헤브론 Braf 저해제를 이용한 암 치료용 조합 요법
US20250064939A1 (en) 2023-07-24 2025-02-27 Sanegene Bio Usa Inc. Lipid-based enhancement agent for rna delivery and therapy
IL326321A (en) 2023-08-04 2026-04-01 Alnylam Pharmaceuticals Inc Methods and compositions for treating CTNNB1-related disorders
WO2025052278A1 (fr) 2023-09-05 2025-03-13 Genevant Sciences Gmbh Lipides cationiques à base de pyrrolidine pour administration de nanoparticules lipidiques d'agents thérapeutiques à des cellules stellaires hépatiques
IL326940A (en) 2023-09-21 2026-05-01 Alnylam Pharmaceuticals Inc Activin A Receptor Type 1C (ACVR1C) IRNA Compositions and Methods of Using Them
WO2025072672A2 (fr) 2023-09-27 2025-04-03 Judo Bio, Inc. Agents d'acides nucléiques modulateurs ciblant slc6a19
WO2025072713A1 (fr) 2023-09-27 2025-04-03 Judo Bio, Inc. Polymyxines pour l'administration d'agents au rein
AU2024348221A1 (en) 2023-09-27 2026-04-30 Judo Bio Inc. Aminoglycosides for delivery of agents to the kidney
WO2025076031A2 (fr) 2023-10-03 2025-04-10 Alnylam Pharmaceuticals, Inc. Macrophages péritonéaux comprenant une nanoparticule encapsulant une molécule d'acide nucléique et leurs méthodes d'utilisation
WO2025090427A1 (fr) 2023-10-23 2025-05-01 University Of Rochester Soulagement d'hyperexcitabilité ciblé glial dans des maladies neurodégénératives
WO2025128799A1 (fr) 2023-12-12 2025-06-19 Korro Bio, Inc. Oligonucléotides d'édition d'arn double brin et leurs utilisations
WO2025133951A1 (fr) 2023-12-21 2025-06-26 Genevant Sciences Gmbh Lipides ionisables appropriés pour des nanoparticules lipidiques
WO2025172510A1 (fr) 2024-02-15 2025-08-21 Fundació Institut D'investigació Biomèdica De Bellvitge (Idibell) Composés pour le traitement de maladies
EP4616864A1 (fr) 2024-03-11 2025-09-17 Peptomyc, S.L. Composé pour le traitement de maladies auto-immunes
WO2025255388A1 (fr) 2024-06-05 2025-12-11 Camp4 Therapeutics Corporation Modulation de la transcription du gène syngap1 à l'aide d'oligonucléotides antisens ciblant les arn régulateurs
WO2026006436A1 (fr) 2024-06-25 2026-01-02 Korro Bio, Inc. Procédés et compositions pour l'édition médiée par adar d'une protéine de liaison à l'adn tar 43 kda (tdp43)
WO2026050125A1 (fr) 2024-08-28 2026-03-05 Alnylam Pharmaceuticals, Inc. Procédés et compositions pour le traitement d'un trouble associé à l'angiotensinogène (agt)
WO2026085164A2 (fr) 2024-10-16 2026-04-23 Alnylam Pharmaceuticals, Inc. Formulations d'arni de snca et leurs procédés d'utilisation

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5168053A (en) * 1989-03-24 1992-12-01 Yale University Cleavage of targeted RNA by RNAase P
US5272263A (en) * 1989-04-28 1993-12-21 Biogen, Inc. DNA sequences encoding vascular cell adhesion molecules (VCAMS)
IE911115A1 (en) * 1990-04-10 1991-10-23 Canji Inc Gene therapy for cell proliferative diseases
GB9106678D0 (en) * 1991-03-28 1991-05-15 Ferguson Mark W J Wound healing
ZA938208B (en) * 1992-11-03 1995-01-03 Gene Shears Pty Ltd TNF-alpha ribozymes and degradation resistant mRNA derivatives linked to TNF-alpha ribozymes
EP0746614A1 (fr) * 1994-02-23 1996-12-11 Ribozyme Pharmaceuticals, Inc. Procede et reactif inhibiteur de l'expression de genes concernant une affection

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AU4769893A (en) 1994-02-14
EP0786522A3 (fr) 1997-08-27
EP1251170A2 (fr) 2002-10-23
AU5209698A (en) 1998-03-19
WO1994002595A1 (fr) 1994-02-03
MX9304329A (es) 1994-05-31
EP1251170A3 (fr) 2002-10-30
EP0654077A4 (fr) 1996-03-13
CA2140343A1 (fr) 1994-02-03
EP0654077A1 (fr) 1995-05-24
JPH07509133A (ja) 1995-10-12

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